uu.seUppsala universitets publikationer
Ändra sökning
Avgränsa sökresultatet
1 - 16 av 16
RefereraExporteraLänk till träfflistan
Permanent länk
Referera
Referensformat
  • apa
  • ieee
  • modern-language-association
  • vancouver
  • Annat format
Fler format
Språk
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Annat språk
Fler språk
Utmatningsformat
  • html
  • text
  • asciidoc
  • rtf
Träffar per sida
  • 5
  • 10
  • 20
  • 50
  • 100
  • 250
Sortering
  • Standard (Relevans)
  • Författare A-Ö
  • Författare Ö-A
  • Titel A-Ö
  • Titel Ö-A
  • Publikationstyp A-Ö
  • Publikationstyp Ö-A
  • Äldst först
  • Nyast först
  • Skapad (Äldst först)
  • Skapad (Nyast först)
  • Senast uppdaterad (Äldst först)
  • Senast uppdaterad (Nyast först)
  • Disputationsdatum (tidigaste först)
  • Disputationsdatum (senaste först)
  • Standard (Relevans)
  • Författare A-Ö
  • Författare Ö-A
  • Titel A-Ö
  • Titel Ö-A
  • Publikationstyp A-Ö
  • Publikationstyp Ö-A
  • Äldst först
  • Nyast först
  • Skapad (Äldst först)
  • Skapad (Nyast först)
  • Senast uppdaterad (Äldst först)
  • Senast uppdaterad (Nyast först)
  • Disputationsdatum (tidigaste först)
  • Disputationsdatum (senaste först)
Markera
Maxantalet träffar du kan exportera från sökgränssnittet är 250. Vid större uttag använd dig av utsökningar.
  • 1.
    Anderson, Henrik
    Uppsala universitet, Teknisk-naturvetenskapliga vetenskapsområdet, Tekniska sektionen, Institutionen för teknikvetenskaper, Fasta tillståndets elektronik.
    Development of Electroacoustic Sensors for Biomolecular Interaction Analysis2009Doktorsavhandling, sammanläggning (Övrigt vetenskapligt)
    Abstract [en]

    Biomolecular interaction analysis to determine the kinetics and affinity between interacting partners is important for the fundamental understanding of biology, as well as for the development of new pharmaceutical substances. A quartz crystal microbalance instrument suitable for kinetics and affinity analyses of interaction events was developed. The functionality of the sensor system was demonstrated by development of an assay for relative affinity determination of lectin-carbohydrate interactions.

    Sensor surfaces allowing for effective immobilization of one interacting partner is a key functionality of a biosensor. Here, three different surfaces and immobilization methods were studied. First, optimized preparation conditions for sensor surfaces based on carboxyl-terminated self assembled monolayers were developed and were demonstrated to provide highly functional biosensor surfaces with low non-specific binding. Second, a method allowing for immobilization of very acidic biomolecules based on the use of an electric field was developed and evaluated. The electric field made it possible to immobilize the highly acidic C-peptide on a carboxylated surface. Third, a method for antibody immobilization on a carboxyl surface was optimized and the influence of immobilization pH on the immobilization level and antigen binding capacity was thoroughly assessed. The method showed high reproducibility for a set of antibodies and allowed for antibody immobilization also at low pH.

    Three broadly different strategies to increase the sensitivity of electroacoustic sensors were explored. A QCM sensor with small resonator electrodes and reduced flow cell dimensions was demonstrated to improve the mass transport rate to the sensor surface. The use of polymers on QCM sensor surfaces to enhance the sensor response was shown to increase the response of an antibody-antigen model system more than ten-fold. Moreover, the application of high frequency thin film bulk acoustic resonators for biosensing was evaluated with respect to sensing range from the surface. The linear detection range of the thin film resonator was determined to be more than sufficient for biosensor applications involving, for instance, antibody-antigen interactions. Finally, a setup for combined frequency and resistance measurements was developed and was found to provide time resolved data suitable for kinetics determination.

    Delarbeten
    1. Study of real-time lectin-carbohydrate interactions on the surface of a quartz crystal microbalance
    Öppna denna publikation i ny flik eller fönster >>Study of real-time lectin-carbohydrate interactions on the surface of a quartz crystal microbalance
    2005 (Engelska)Ingår i: Biosensors & Bioelectronics, Vol. 21, s. 60-66Artikel i tidskrift (Refereegranskat) Published
    Identifikatorer
    urn:nbn:se:uu:diva-26504 (URN)
    Tillgänglig från: 2007-02-19 Skapad: 2007-02-19 Senast uppdaterad: 2011-01-12
    2. Esterification of Self-Assembled Carboxylic-Acid-Terminated Thiol Monolayers in Acid Environment: A Time-Dependent Study
    Öppna denna publikation i ny flik eller fönster >>Esterification of Self-Assembled Carboxylic-Acid-Terminated Thiol Monolayers in Acid Environment: A Time-Dependent Study
    Visa övriga...
    2010 (Engelska)Ingår i: Langmuir, ISSN 0743-7463, E-ISSN 1520-5827, Vol. 26, nr 2, s. 821-829Artikel i tidskrift (Refereegranskat) Published
    Abstract [en]

    This contribution reports on the influence of acids oil the quality of carboxylic-acid-terminated self-assembled monolayers (SAMs) on gold prepared from ethanolic solution of HS-(CH2)(15)-COOH and HS-(CH2)(11)CONH-(EG)(6)CH2-COOH. Null ellipsometry, contact angle goniometry, and infrared reflection-absorption spectroscopy are used to monitor the physical and chemical changes occurring within the SAMs upon acid post treatment; after incubation with acids present in the solution: and after incubation in aged acid containing solutions. The presence of acid has a positive effect oil the crystallinity, packing, and orientation Of the Supporting alkyl and ethylene glycol Subunits of the SAM. Our Studies also confirm previous findings stating that the carboxylic groups are rapidly converted into ethyl ester groups in the presence of hydrochloric acid in the incubation solution. It is also evident that the conversion occurs in the presence of the weaker acid, acetic acid, although at a much slower rate than that for hydrochloric acid. This is a new observation that has not been reported on before. The physical and chemical characterization is also complemented with a functional bioaffinity study. The functional evaluation revealed that the present model system was surprisingly insensitive to the degree of esterification of the carboxylic acid groups, but that 4 weeks of storage of the two investigated thiols in hydrochloric acid containing ethanol resulted in SAMs that were completely inactive with respect to immobilization and subsequent binding of the antigen. It was encouraging to note that the nonspecific binding of both antigen and antibody was extremely low oil the two SAMs, regardless of the relative amount of ethyl esters on the surface.

    Nationell ämneskategori
    Teknik och teknologier Naturvetenskap
    Identifikatorer
    urn:nbn:se:uu:diva-137945 (URN)10.1021/la902255j (DOI)000273403400029 ()
    Tillgänglig från: 2010-12-16 Skapad: 2010-12-16 Senast uppdaterad: 2017-12-11Bibliografiskt granskad
    3. Electroimmobilization of proinsulin C-peptide to a quartz crystal microbalance sensor chip for protein affinity purification
    Öppna denna publikation i ny flik eller fönster >>Electroimmobilization of proinsulin C-peptide to a quartz crystal microbalance sensor chip for protein affinity purification
    Visa övriga...
    2005 (Engelska)Ingår i: Anal Biochem, Vol. 341, nr 1, s. 89-93Artikel i tidskrift (Refereegranskat) Published
    Identifikatorer
    urn:nbn:se:uu:diva-26506 (URN)
    Tillgänglig från: 2007-02-19 Skapad: 2007-02-19 Senast uppdaterad: 2011-01-12
    4. Optimizing immobilization on two-dimensional carboxyl surface: pH dependence of antibody orientation and antigen binding capacity
    Öppna denna publikation i ny flik eller fönster >>Optimizing immobilization on two-dimensional carboxyl surface: pH dependence of antibody orientation and antigen binding capacity
    Visa övriga...
    2010 (Engelska)Ingår i: Analytical Biochemistry, ISSN 0003-2697, E-ISSN 1096-0309, Vol. 398, nr 2, s. 161-168Artikel i tidskrift (Refereegranskat) Published
    Abstract [en]

    The performance of immunosensors is highly dependent on the amount of immobilized antibodies and their remaining antigen binding capacity. In this work, a method for immobilization of antibodies on a two dimensional carboxyl surface has been optimized using quartz crystal microbalance biosensors. We have shown that successful immobilization is highly dependent on surface pKa, antibody pI and pH of immobilization buffer. By use of EDC/sulfo-NHS activation reagents, the effect of the intrinsic surface pKa is avoided and immobilization also at very low pH has been made possible which is of importance for immobilization of acidic proteins. Generic immobilization conditions were demonstrated on a panel of antibodies which resulted in an average coefficient of variation of 4% for the immobilization of these antibodies.

    Antigen binding capacity as a function of immobilization pH was studied. In most cases the antigen binding capacity followed the immobilization response. However, the antigen to antibody binding ratio differed between the antibodies investigated, and for one of the antibodies, the antigen binding capacity was significantly lower than expected from immobilization in a certain pH range. Tests with anti-Fc and anti-Fab antibodies on different antibody surfaces showed that the orientation of the antibodies on the surface had a profound effect on the antigen binding capacity of the immobilized antibodies.

    Nyckelord
    antibody, orientation, biosensor, immobilization, sensor surface, QCM
    Nationell ämneskategori
    Annan industriell bioteknik Analytisk kemi
    Forskningsämne
    Ytbioteknik
    Identifikatorer
    urn:nbn:se:uu:diva-107233 (URN)10.1016/j.ab.2009.11.038 (DOI)000274615100003 ()19962366 (PubMedID)
    Tillgänglig från: 2009-07-30 Skapad: 2009-07-30 Senast uppdaterad: 2017-12-13Bibliografiskt granskad
    5. Quartz crystal microbalance sensor design: I. Experimental study of sensor response and performance
    Öppna denna publikation i ny flik eller fönster >>Quartz crystal microbalance sensor design: I. Experimental study of sensor response and performance
    Visa övriga...
    2007 (Engelska)Ingår i: Sensors and actuators. B, Chemical, ISSN 0925-4005, E-ISSN 1873-3077, Vol. 123, nr 1, s. 27-34Artikel i tidskrift (Refereegranskat) Published
    Abstract [en]

    This paper investigates a novel quartz crystal microbalance (QCM) biosensor with a small and rectangular flow cell along with a correspondingly shaped crystal electrode. The sensor was evaluated with impedance analysis and compared to standard circular sensor crystals and sensor crystals with small circular electrodes. Comparative QCM measurements on an antibody–antigen interaction system were carried out on the rectangular and standard circular sensor systems. Impedance analysis and subsequent data extraction of the three different sensor crystals showed that the smaller sensors had significantly higher Q-values in air, but that liquid load on the electrodes lowered the Q-values radically for all crystals. Under liquid load, Q-values for the standard circular and the rectangular sensors were similar whereas the Q-value for the small circular sensor was 50% higher. QCM experiments showed that the QCM system with rectangular crystal electrodes was fully functional in a liquid environment. The rectangular system showed higher and more rapid responses for series of antibody injections, albeit at a higher noise level than the standard system. The study elucidates a significant potential for improvement of sensor performance by optimising the sensor electrode size and shape together with the flow cell geometry.

    Nyckelord
    QCM, Biosensor, Sensitivity, Mass transport, Protein interactions
    Nationell ämneskategori
    Teknik och teknologier
    Identifikatorer
    urn:nbn:se:uu:diva-94271 (URN)10.1016/j.snb.2006.07.027 (DOI)000246171200006 ()
    Tillgänglig från: 2006-04-07 Skapad: 2006-04-07 Senast uppdaterad: 2017-12-14Bibliografiskt granskad
    6. Quartz crystal microbalance biosensor design: II. Simulation of sample transport
    Öppna denna publikation i ny flik eller fönster >>Quartz crystal microbalance biosensor design: II. Simulation of sample transport
    2007 (Engelska)Ingår i: Sensors and actuators. B, Chemical, ISSN 0925-4005, E-ISSN 1873-3077, Vol. 123, nr 1, s. 21-26Artikel i tidskrift (Refereegranskat) Published
    Abstract [en]

    The influence of flow cell geometry on sample dispersion in a quartz crystal microbalance (QCM) biosensor system was investigated. A circular and a rectangular flow cell and corresponding sensor electrodes were studied experimentally and modelled using a coupled Navier-Stokes and convection-diffusion model. Finite element simulations showed that dispersion phenomena in a flow cell can be significantly reduced with the rectangular flow cell compared to a circular system. Experimental results from measurement of the time-dependent viscosity change of a model sample indicate that the sample delivery system has a predominant effect on the dispersion of the whole sensor system. Consequently, improvement of the sensor flow cell should be accompanied with improvement of the sample delivery system. With reference to kinetic studies of biological interactions, the current dispersion should have little effect on the results for studies of interaction pairs with relatively slow to normal binding rates such as antibody-antigen interactions. Incentive for further development of the flow cell and sample delivery system exists primarily for applications with high reaction rates such as for certain receptor ligand interactions.

     

    Nyckelord
    QCM, Simulation, FEM, Navier–Stokes, Convection and diffusion, Microfluidic
    Nationell ämneskategori
    Teknik och teknologier
    Identifikatorer
    urn:nbn:se:uu:diva-94272 (URN)10.1016/j.snb.2006.07.028 (DOI)000246171200005 ()
    Tillgänglig från: 2006-04-07 Skapad: 2006-04-07 Senast uppdaterad: 2017-12-14Bibliografiskt granskad
    7. Surface-Confined Photopolymerization of pH-Responsive Acrylamide/Acrylate Brushes on Polymer Thin Films
    Öppna denna publikation i ny flik eller fönster >>Surface-Confined Photopolymerization of pH-Responsive Acrylamide/Acrylate Brushes on Polymer Thin Films
    Visa övriga...
    2008 (Engelska)Ingår i: Langmuir, ISSN 0743-7463, E-ISSN 1520-5827, Vol. 24, nr 14, s. 7559-7564Artikel i tidskrift (Refereegranskat) Published
    Abstract [en]

    Dynamic acrylamide/acrylate polymeric brushes were synthesized at gold-plated quartz crystal surfaces. The crystals were initially coated with polystyrene-type thin films, derivatized with photolabile iniferter groups, and subsequently subjected to photoinitiated polymerization in acrylamide/acrylate monomer feeds. This surface-confined polymerizationmethod enabled direct photocontrol over the polymerization, as followed by increased frequency responses of the crystal oscillations in a quartz crystal microbalance (QCM). The produced polymer layers were also found to be highlysensitive to external acid/base stimuli. Large oscillation frequency shifts were detected when the brushes were exposedto buffer solutions of different pH. The dynamic behavior of the resulting polymeric brushes was evaluated, and theextent of expansion and contraction of the films was monitored by the QCM setup in situ in real time. The resultingresponses were rapid, and the effects were fully reversible. Low pH resulted in full contractions of the films, whereashigher pH yielded maximal expansion in order to minimize repulsion around the charged acrylate centers. The surfacesalso proved to be very robust because the responsiveness was reproducible over many cycles of repeated expansionand contraction. Using ellipsometry, copolymer layers were estimated to be ∼220 nm in a collapsed state and ∼340nm in the expanded state, effectively increasing the thickness of the film by 55%.

    Nyckelord
    Surface grafting, polymer brushes, hydrogels, sensor surfaces, transfer radical polymerization
    Nationell ämneskategori
    Polymerkemi Fysikalisk kemi Teknik och teknologier
    Forskningsämne
    Ytbioteknik
    Identifikatorer
    urn:nbn:se:uu:diva-107234 (URN)10.1021/la800700h (DOI)000257468300072 ()18563922 (PubMedID)
    Tillgänglig från: 2009-07-30 Skapad: 2009-07-30 Senast uppdaterad: 2017-12-13Bibliografiskt granskad
    8. Signal Enhancement in Ligand-Receptor Interactions using Dynamic Polymers at Quartz Crystal Microbalance Surfaces
    Öppna denna publikation i ny flik eller fönster >>Signal Enhancement in Ligand-Receptor Interactions using Dynamic Polymers at Quartz Crystal Microbalance Surfaces
    Visa övriga...
    2016 (Engelska)Ingår i: The Analyst, ISSN 0003-2654, E-ISSN 1364-5528, Vol. 141, nr 13, s. 3993-3996Artikel i tidskrift (Refereegranskat) Published
    Abstract [en]

    The potential for signal amplification on QCM sensors by use of in situ polymerized poly(acrylic acid) brushes has been studied. A biotin derivative was immobilized on these surfaces and the interaction with anti-biotin Fabs was evaluated. Interaction data was found to be specific for the studied binding events, and the level of non-specific binding was shown to be low. The surface was proven to be suitable for regeneration, of importance for biomolecular interaction analysis and repetitive immunoassays.

    For comparison, the same interaction system was tested using commercial sensor surfaces with carboxylated self-assembled monolayers. The poly(acrylic acid) surface showed a dramatic increase in signal response with more than ten times the signal of the carboxylated self-assembled monolayer surface. Thus, the present study shows that polymers can be successfully applied to amplify responses on QCM sensors, valuable for studies of interactions between receptors and low molecular weight compounds.

    Nyckelord
    dynamic chemistry, quartz crystal microbalance, iniferter, photopolymerization, biosensor, interaction
    Nationell ämneskategori
    Polymerkemi Teknik och teknologier
    Forskningsämne
    Ytbioteknik
    Identifikatorer
    urn:nbn:se:uu:diva-107252 (URN)10.1039/c6an00735j (DOI)000378942300006 ()27196531 (PubMedID)
    Externt samarbete:
    Forskningsfinansiär
    Vetenskapsrådet
    Tillgänglig från: 2009-07-30 Skapad: 2009-07-30 Senast uppdaterad: 2017-12-13Bibliografiskt granskad
    9. On the applicability of high frequency acoustic shear mode biosensing in view of thickness limitations set by the film resonance
    Öppna denna publikation i ny flik eller fönster >>On the applicability of high frequency acoustic shear mode biosensing in view of thickness limitations set by the film resonance
    Visa övriga...
    2009 (Engelska)Ingår i: Biosensors & bioelectronics, ISSN 0956-5663, E-ISSN 1873-4235, Vol. 24, nr 11, s. 3387-3390Artikel i tidskrift (Refereegranskat) Published
    Abstract [en]

    The IC-compatible thin film bulk acoustic resonator (FBAR) technology has made it possible to move the thickness excited shear mode sensing of biological layers into a new sensing regime using substantially higher operation frequencies than the conventionally used Quartz Crystal Microbalance (QCM). The limitations of the linear range set by the film resonance using viscoelastic protein films are here for the first time addressed specifically for FBARs operating at 700MHz up to 1.5GHz. Two types of protein multilayer sensing were employed; one utilizing alternating layers of Streptavidin and Biotinated BSA and the other using stepwise cross-linking of fibrinogen with EDC/NHS activation of its carboxyl groups. In both cases the number of protein layers within the linear regime is well above the number of protein layers usually used in biosensor applications, further verifying the applicability of the FBAR as a biosensor. Theoretical calculations are also presented using well established physical models to illustrate the expected behavior of the FBAR sensor, in view of both the frequency and the dissipation shifts.

    Nyckelord
    Shear mode electroacoustic sensing, Streptavidin–biotinated BSA, Fibrinogen, High frequency, Thin film bulk acoustic resonator (FBAR)
    Nationell ämneskategori
    Teknik och teknologier
    Forskningsämne
    Elektronik
    Identifikatorer
    urn:nbn:se:uu:diva-89406 (URN)10.1016/j.bios.2009.04.021 (DOI)000267577900035 ()
    Tillgänglig från: 2009-02-12 Skapad: 2009-02-12 Senast uppdaterad: 2018-06-26Bibliografiskt granskad
    10. Systematic investigation of biomolecular interactions using combined frequency and motional resistance measurements
    Öppna denna publikation i ny flik eller fönster >>Systematic investigation of biomolecular interactions using combined frequency and motional resistance measurements
    Visa övriga...
    2011 (Engelska)Ingår i: Sensors and actuators. B, Chemical, ISSN 0925-4005, E-ISSN 1873-3077, Vol. 153, nr 1, s. 135-144Artikel i tidskrift (Refereegranskat) Published
    Abstract [en]

    The resonance frequency of acoustic biosensors is today used as a label-free technique for detecting mass changes on sensor surfaces. In combination with an appropriate continuous flow system it has earlier been used for affinity and kinetic rate determination. Here, we assess the potential of a modified acoustic biosensor, monitoring also the real-time dissipation through the resistance of the sensor, to obtain additional kinetic information related to the structure and conformation of the molecules on the surface. Actual interaction studies, including an attempt to determine avidity, are presented along with thorough verification of the experimental setup utilizing true viscous load exposure together with protein and DNA immobilizations. True viscous loads show a linear relationship between resistance and frequency as expected. However, in the interaction studies between antibodies and proteins, as well as in the immobilization of DNA and proteins, higher surface concentrations of interacting molecules led to a decrease (i.e. deviation from the linear trend) in the differential resistance to frequency ratio. This is interpreted as increased surface rigidity at higher surface concentrations of immobilized molecules. Consequently, studies that aim at obtaining biological binding information, such as avidity, from real-time resistance and dissipation data should be conducted at low surface concentrations. In addition, the differential resistance to frequency relationship was found to be highly dependent on the rigidity of the preceding layer(s) of immobilized molecules. This dependence can be utilized to obtain a higher signal-to-noise ratio for resistance measurement by using low surface densities of immobilized interaction partners.

    Nyckelord
    biosensor, interaction analysis, QCM, dissipation, motional resistance, kinetics
    Nationell ämneskategori
    Analytisk kemi Annan industriell bioteknik
    Forskningsämne
    Analytisk kemi; Teknisk fysik med inriktning mot mikrosystemteknik; Teknisk fysik med inriktning mot elektronik
    Identifikatorer
    urn:nbn:se:uu:diva-107253 (URN)10.1016/j.snb.2010.10.019 (DOI)000289019300020 ()
    Tillgänglig från: 2009-07-30 Skapad: 2009-07-30 Senast uppdaterad: 2018-06-26Bibliografiskt granskad
  • 2.
    Anderson, Henrik
    et al.
    Uppsala universitet, Teknisk-naturvetenskapliga vetenskapsområdet, Tekniska sektionen, Institutionen för teknikvetenskaper, Fasta tillståndets elektronik.
    Wingqvist, Gunilla
    Uppsala universitet, Teknisk-naturvetenskapliga vetenskapsområdet, Tekniska sektionen, Institutionen för teknikvetenskaper, Fasta tillståndets elektronik.
    Weissbach, Thomas
    Attana AB, Stockholm.
    Wallinder, Daniel
    Attana AB, Stockholm.
    Katardjiev, Ilia
    Uppsala universitet, Teknisk-naturvetenskapliga vetenskapsområdet, Tekniska sektionen, Institutionen för teknikvetenskaper, Fasta tillståndets elektronik.
    Ingemarsson, Björn
    Attana AB, Stockholm.
    Systematic investigation of biomolecular interactions using combined frequency and motional resistance measurements2011Ingår i: Sensors and actuators. B, Chemical, ISSN 0925-4005, E-ISSN 1873-3077, Vol. 153, nr 1, s. 135-144Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    The resonance frequency of acoustic biosensors is today used as a label-free technique for detecting mass changes on sensor surfaces. In combination with an appropriate continuous flow system it has earlier been used for affinity and kinetic rate determination. Here, we assess the potential of a modified acoustic biosensor, monitoring also the real-time dissipation through the resistance of the sensor, to obtain additional kinetic information related to the structure and conformation of the molecules on the surface. Actual interaction studies, including an attempt to determine avidity, are presented along with thorough verification of the experimental setup utilizing true viscous load exposure together with protein and DNA immobilizations. True viscous loads show a linear relationship between resistance and frequency as expected. However, in the interaction studies between antibodies and proteins, as well as in the immobilization of DNA and proteins, higher surface concentrations of interacting molecules led to a decrease (i.e. deviation from the linear trend) in the differential resistance to frequency ratio. This is interpreted as increased surface rigidity at higher surface concentrations of immobilized molecules. Consequently, studies that aim at obtaining biological binding information, such as avidity, from real-time resistance and dissipation data should be conducted at low surface concentrations. In addition, the differential resistance to frequency relationship was found to be highly dependent on the rigidity of the preceding layer(s) of immobilized molecules. This dependence can be utilized to obtain a higher signal-to-noise ratio for resistance measurement by using low surface densities of immobilized interaction partners.

  • 3.
    Andersson Schönn, Mikael
    et al.
    Uppsala universitet, Teknisk-naturvetenskapliga vetenskapsområdet, Biologiska sektionen, Institutionen för biologisk grundutbildning.
    Borg, Malin
    Uppsala universitet, Teknisk-naturvetenskapliga vetenskapsområdet, Biologiska sektionen, Institutionen för biologisk grundutbildning.
    Bunpuckdee, Benja
    Uppsala universitet, Teknisk-naturvetenskapliga vetenskapsområdet, Biologiska sektionen, Institutionen för biologisk grundutbildning.
    Gioeli, Erik
    Uppsala universitet, Teknisk-naturvetenskapliga vetenskapsområdet, Biologiska sektionen, Institutionen för biologisk grundutbildning.
    Holdar, Karl
    Uppsala universitet, Teknisk-naturvetenskapliga vetenskapsområdet, Biologiska sektionen, Institutionen för biologisk grundutbildning.
    Odlingsmedium: Att ersätta fetalt kalvserum med ett kemiskt definierat substitut2014Självständigt arbete på grundnivå (kandidatexamen), 10 poäng / 15 hpStudentuppsats (Examensarbete)
    Abstract [sv]

    SammanfattningProjektgrupp 14-X5 avser med denna rapport att ge avdelningen Bioreagens på Thermo Fisher Scientific ett underlag för att på sikt kunna byta ut fetalt kalvserum (FCS) mot ett kemiskt definierat suplement vid odling av mushybridomceller för produktion av monoklonala antikroppar. Thermo Fisher Scientific är ett världsomspännande bioteknikföretag som utvecklar blodtestsystem som stöd för klinisk diagnos och uppföljning av allergier, astma och autoimmuna sjukdomar.

    Fetalt kalvserum är en tillsats i många odlingsmedier som ofta är nödvändig för att cellerna ska växa. Det finns dock många problem med FCS. Det är en biprodukt av köttindustrin och produceras på ett etiskt tveksamt sätt, variation mellan olika batcher förekommer och då det är en animalisk produkt finns en risk för kontamination av bland annat bakterier, virus och prioner. Av dessa anledningar vill man byta ut FCS mot ett kemiskt definierat, serumfritt supplement. Vi har utrett vilka ämnen eller grupper av ämnen som har störst potential att vara bra substitut för FCS, samt rangordnat dessa. Genom våra artikelstudier har vi kommit fram till att man kan dela in alternativen i tre grupper: lipider, tillväxtfaktorer och små biomolekyler. Bland lipiderna är det linol- och oljesyra som i flera artiklar har visats ha god effekt på både celltillväxt och antikroppsproduktion. Kolesterol har även visats ha positiva effekter. Tillväxtfaktorerna som har valts är epidermal growth factor (EGF), fibroblast growth factor (FGF), interleukin-2 (IL-2) och interleukin-6 (IL-6). Dessa har främst en positiv effekt på cellernas antikroppsproduktion. Bland de små biomolekyler som har valts ut finns en mängd olika ämnen som på olika sätt kan bidra till att skapa en bra miljö för hybridomceller i ett serumfritt medium.

    För att få en klarare bild av de olika förslagen har de jämförts. Vi menar att de ämnen som är komponenter i det basala mediet Ham F-12 bör prioriteras då det används som standard vid odling av en bred grupp av celltyper. Denna grupp inkluderar linolsyra, putrescin, tymidin, hypoxantin samt liponsyra. Därefter anser vi att återstående lipider (oljesyra och kolesterol) ska prioriteras då de har visats kunna öka både hybridomcellers tillväxt och antikroppsproduktion i ett serumfritt medium. Tillväxtfaktorer, som främst ökar antikroppsproduktionen hos cellerna  och i vissa fall även deras livslängd placerar vi på tredje plats i vår rangordning. Övriga ämnen i gruppen “Små biomolekyler” (paraaminobensoesyra och glutation) prioriterar vi sist då de inte är komponenter i Ham F-12 och de inte explicit visats påverka celltillväxten ellerantikroppsproduktionen, men har identifierats i det serumfria mediet IBL media III.

  • 4.
    Barkenäs, Emelie
    Uppsala universitet, Teknisk-naturvetenskapliga vetenskapsområdet, Biologiska sektionen, Institutionen för biologisk grundutbildning.
    Automation of a solid-phase proximity ligation assay for biodefense applications2013Självständigt arbete på avancerad nivå (yrkesexamen), 20 poäng / 30 hpStudentuppsats (Examensarbete)
    Abstract [en]

    The extent of devastation caused by a biological warfare attack is highly correlated to the time from release to detection. As a step towards lowering the detection time the international project TWOBIAS was launched. Here, the main goal is to develop an automated, specific and sensitive combined detection and identification instrument capable of identifying a biological threat within an hour. The identification unit is comprised of a sample preparation module, an amplification module and a detection module and utilizes a proximity ligation assay in combination with circle-to-circle amplification in order to detect a biological threat. This thesis describes the automation of the sample preparation steps of the assay and the integration with the downstream units. The functionality of the sample preparation module was verified by subjecting it to biological samples in a laboratory and at a real-life location. The results showed that the sample preparation module was capable of preparing a sample collected in a complex environment with the same results as a sample prepared in a laboratory. 

  • 5.
    Caldwell, Karin D.
    et al.
    Uppsala universitet, Teknisk-naturvetenskapliga vetenskapsområdet, Kemiska sektionen, Institutionen för fysikalisk och analytisk kemi.
    Janson, Jan-Christer
    Uppsala universitet, Teknisk-naturvetenskapliga vetenskapsområdet, Kemiska sektionen, Institutionen för fysikalisk och analytisk kemi.
    The Origin of Sepahdex2006Ingår i: GIT Laboratory Journal: Europe, Vol. 10, nr 5, s. 18-20Artikel i tidskrift (Övrigt vetenskapligt)
  • 6.
    Cheng, Wing-Shing
    Uppsala universitet, Medicinska vetenskapsområdet, Medicinska fakulteten, Institutionen för onkologi, radiologi och klinisk immunologi, Enheten för klinisk immunologi.
    TARP Promoter-Based Prostate Cancer Gene Therapy: From Development to Application2005Doktorsavhandling, sammanläggning (Övrigt vetenskapligt)
    Abstract [en]

    Prostate cancer is one leading cause of cancer-related death among men in Western countries. The standard therapies for localized prostate cancer include radical prostatectomy and radiation therapy. Such measures are relatively effective in the short term, but many patients ultimately relapse. These patients may benefit from a combination of standard therapy and oncolytic virus therapy. My work aimed to develop viruses for this purpose.

    TARP is a protein that in males is specifically expressed in prostate epithelial and cancer cells. In my thesis, I characterized the TARP promoter and showed that TARP expression is regulated at the transcriptional level by testosterone through binding of the androgen receptor in the proximal TARP promoter. I further developed TARP promoter-based regulatory sequences for prostate-specific gene expression. A sequence comprising a PSA enhancer, a PSMA enhancer and the TARP promoter was constructed and designated PPT. An adenoviral vector containing the PPT sequence shielded from transcriptional interference by an H19 insulator showed high prostate-specific transcriptional activity in human cells both in the presence and absence of testosterone. However, in experimental murine prostate cancer the PPT sequence is not active. Therefore, a two-step transcriptional amplification (TSTA) system was used together with the PPT sequence to develop an adenovirus that confers prostate-specific transgene expression also in murine cells.

    I constructed a conditionally replicating adenovirus where the E1A gene expression is controlled by an H19 insulator-shielded PPT regulatory sequence, Ad[I/PPT-E1A]. This virus exhibited absolute prostate specificity in terms of E1A expression, viral replication and cytolysis in vitro and in vivo. Importantly, our virus is active both in the presence and absence of testosterone, which may prove beneficial for patients treated by hormonal withdrawal.

    Hopefully, my work will improve existing gene therapy strategies for prostate cancer and in the long term improve the prognosis for patients with prostate cancer.

    Delarbeten
    1. Characterization of the Androgen-Regulated Prostate-Specific TARP Promoter
    Öppna denna publikation i ny flik eller fönster >>Characterization of the Androgen-Regulated Prostate-Specific TARP Promoter
    2003 Ingår i: Endocrinology, ISSN 0013-7227, Vol. 144, nr 8, s. 3433-3440Artikel i tidskrift (Refereegranskat) Published
    Identifikatorer
    urn:nbn:se:uu:diva-92860 (URN)
    Tillgänglig från: 2005-04-11 Skapad: 2005-04-11Bibliografiskt granskad
    2. A novel TARP-promoter-based adenovirus against hormone-dependent and hormone-refractory prostate cancer
    Öppna denna publikation i ny flik eller fönster >>A novel TARP-promoter-based adenovirus against hormone-dependent and hormone-refractory prostate cancer
    Visa övriga...
    2004 (Engelska)Ingår i: Molecular Therapy, ISSN 1525-0016, E-ISSN 1525-0024, Vol. 10, nr 2, s. 355-364Artikel i tidskrift (Refereegranskat) Published
    Abstract [en]

    TARP (T cell receptor gamma-chain alternate reading frame protein) is a protein that in males is uniquely expressed in prostate epithelial cells and prostate cancer cells. We have previously shown that the transcriptional activity of a chimeric sequence comprising the TARP promoter (TARPp) and the PSA enhancer (PSAe) is strictly controlled by testosterone and highly restricted to cells of prostate origin. Here we report that a chimeric sequence comprising TARPp and the PSMA enhancer (PSMAe) is highly active in testosterone-deprived prostate cancer cells, while a regulatory sequence comprising PSAe, PSMAe, and TARPp (PPT) has high prostate-specific activity both in the presence and in the absence of testosterone. Therefore, the PPT sequence may, in a gene therapy setting, be beneficial to prostate cancer patients that have been treated with androgen withdrawal. A recombinant adenovirus vector with the PPT sequence, shielded from interfering adenoviral sequences by the mouse H19 insulator, yields high and prostate-specific transgene expression both in cell cultures and when prostate cancer, PC-346C, tumors were grown orthotopically in nude mice. Intravenous virus administration reveals both higher activity and higher selectivity for the insulator-shielded PPT sequence than for the immediate-early CMV promoter. Therefore, we believe that an adenovirus with therapeutic gene expression controlled by an insulator-shielded PPT sequence is a promising candidate for gene therapy of prostate cancer.

    Nyckelord
    Adenoviridae/*genetics, Animals, Cell Line; Tumor, Enhancer Elements (Genetics)/genetics, Gene Expression Regulation; Neoplastic, Gene Therapy/*methods, Genes; Reporter/genetics, Genetic Vectors/genetics, Humans, Insulator Elements/genetics, Luciferases/analysis/genetics, Male, Mice, Neoplasms; Hormone-Dependent/genetics/*metabolism/therapy, Nuclear Proteins/*genetics, Promoter Regions (Genetics)/*genetics, Prostatic Neoplasms/genetics/*metabolism/therapy, Research Support; Non-U.S. Gov't, Testosterone/metabolism
    Nationell ämneskategori
    Medicin och hälsovetenskap
    Identifikatorer
    urn:nbn:se:uu:diva-92861 (URN)10.1016/j.ymthe.2004.05.022 (DOI)15294182 (PubMedID)
    Tillgänglig från: 2005-04-11 Skapad: 2005-04-11 Senast uppdaterad: 2017-12-14Bibliografiskt granskad
    3. An oncolytic conditionally replicating adenovirus for hormone-dependent and hormone-independent prostate cancer
    Öppna denna publikation i ny flik eller fönster >>An oncolytic conditionally replicating adenovirus for hormone-dependent and hormone-independent prostate cancer
    Visa övriga...
    2006 (Engelska)Ingår i: Cancer Gene Therapy, ISSN 0929-1903, E-ISSN 1476-5500, Vol. 13, nr 1, s. 13-20Artikel i tidskrift (Refereegranskat) Published
    Abstract [en]

    The use of conditionally replicating adenoviruses offers an attractive complementary treatment strategy for localized prostate cancer. We have produced a replicating adenovirus, Ad[I/PPT-E1A], where E1A gene expression is controlled by a recombinant regulatory sequence designated PPT. The PPT sequence comprises a PSA enhancer, a PSMA enhancer and a T-cell receptor gamma-chain alternate reading frame protein promoter, and it is shielded from transcriptional interference from adenoviral backbone sequences by an H19 insulator. Ad[I/PPT-E1A] yields prostate-specific E1A protein expression, viral replication and cytolysis in vitro. Furthermore, Ad[I/PPT-E1A] considerably regresses the growth of subcutaneous LNCaP prostate cancer tumors in nude mice. Importantly, the viral replication and cytolytic effect of Ad[I/PPT-E1A] are independent of the testosterone levels in the prostate cancer cells. This may be beneficial in a clinical setting since many prostate cancer patients are treated with androgen withdrawal. In conclusion, Ad[I/PPT-E1A] may prove to be useful in the treatment of localized prostate cancer.

    Nyckelord
    Adenoviridae/*metabolism, Adenovirus E1A Proteins/genetics/metabolism, Animals, Gene Expression Regulation; Neoplastic, Genetic Vectors/metabolism/therapeutic use, Humans, Male, Mice, Mice; Inbred C57BL, Mice; Nude, Neoplasms; Hormone-Dependent/genetics/*metabolism, Prostatic Neoplasms/*metabolism, Testosterone/metabolism, Time Factors, Transfection, Virus Replication
    Nationell ämneskategori
    Medicin och hälsovetenskap
    Identifikatorer
    urn:nbn:se:uu:diva-92862 (URN)10.1038/sj.cgt.7700881. (DOI)16052227 (PubMedID)
    Tillgänglig från: 2005-04-11 Skapad: 2005-04-11 Senast uppdaterad: 2017-12-14Bibliografiskt granskad
    4. Two-step transcriptional amplification (TSTA) of the PPT regulatory sequence in murine and human prostate cancer cell lines
    Öppna denna publikation i ny flik eller fönster >>Two-step transcriptional amplification (TSTA) of the PPT regulatory sequence in murine and human prostate cancer cell lines
    Manuskript (Övrigt vetenskapligt)
    Identifikatorer
    urn:nbn:se:uu:diva-92863 (URN)
    Tillgänglig från: 2005-04-11 Skapad: 2005-04-11 Senast uppdaterad: 2010-01-13Bibliografiskt granskad
  • 7.
    Dahl, Fredrik
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för genetik och patologi.
    Selector Technology: For Multiplex DNA Analysis2005Doktorsavhandling, sammanläggning (Övrigt vetenskapligt)
    Abstract [en]

    A majority of methods for identifying sequences in the human genome involve target sequence amplification through PCR. This work presents novel methods for amplifying circularized DNA and presents solutions for some major limitations of PCR.

    We have developed a novel method to amplify circularized DNA molecules based on a serial rolling-circle replication reaction, called circle to circle amplification (C2CA). Amplified DNA circles can be detected in array-based analyses or in real-time using molecular beacons. The amplification mechanism allows higher precision in quantification than in exponential amplification methods like PCR, and more products can be generated than in PCR.

    A major limitation of PCR is that amplification artifacts arise when large numbers of specific primer pairs are simultaneously added to a reaction. We have developed a solution to this problem that enables multiplex PCR amplification of specific target sequences without producing amplification artifacts. The procedure is based on oligonucleotide constructs, called selectors. The selectors identify defined target nucleic acid sequences, and they act as ligation templates to direct circularization of these targets. The selectors contain a general primer-pair motif that allows the circularized targets to be amplified in multiplex using a universal PCR primer pair. We also developed a computer program, PieceMaker, that finds an optimal design of selector probes for a given selector application. We demonstrate the method by performing a 96-plex PCR of specific DNA sequences with high success-rate and reproducibility.

    Delarbeten
    1. Real-time monitoring of rolling-circle amplification using a modified molecular beacon design
    Öppna denna publikation i ny flik eller fönster >>Real-time monitoring of rolling-circle amplification using a modified molecular beacon design
    Visa övriga...
    2002 (Engelska)Ingår i: Nucleic Acids Research, ISSN 0305-1048, E-ISSN 1362-4962, Vol. 30, nr 14, s. e66-Artikel i tidskrift (Refereegranskat) Published
    Abstract [en]

    We describe a method to monitor rolling-circle replication of circular oligonucleotides in dual-color and in real-time using molecular beacons. The method can be used to study the kinetics of the polymerization reaction and to amplify and quantify circularized oligonucleotide probes in a rolling-circle amplification (RCA) reaction. Modified molecular beacons were made of 2'-O-Me-RNA to prevent 3' exonucleolytic degradation by the polymerase used. Moreover, the complement of one of the stem sequences of the molecular beacon was included in the RCA products to avoid fluorescence quenching due to inter-molecular hybridization of neighboring molecular beacons hybridizing to the concatemeric polymerization product. The method allows highly accurate quantification of circularized DNA over a broad concentration range by relating the signal from the test DNA circle to an internal reference DNA circle reporting in a distinct fluorescence color.

    Nationell ämneskategori
    Naturvetenskap
    Identifikatorer
    urn:nbn:se:uu:diva-93438 (URN)10.1093/nar/gnf065 (DOI)12136114 (PubMedID)
    Tillgänglig från: 2005-09-14 Skapad: 2005-09-14 Senast uppdaterad: 2017-12-14Bibliografiskt granskad
    2. Circle-to-circle amplification for precise and sensitive DNA analysis.
    Öppna denna publikation i ny flik eller fönster >>Circle-to-circle amplification for precise and sensitive DNA analysis.
    Visa övriga...
    2004 (Engelska)Ingår i: Proc Natl Acad Sci U S A, ISSN 0027-8424, Vol. 101, nr 13, s. 4548-53Artikel i tidskrift (Övrigt vetenskapligt) Published
    Nyckelord
    Base Sequence, DNA/chemistry/*genetics, DNA Replication/genetics, DNA; Circular/chemistry/*genetics, Gene Amplification, Genetic Diseases; Inborn/genetics, Humans, Models; Molecular, Molecular Sequence Data, Nucleic Acid Conformation, Research Support; Non-U.S. Gov't
    Identifikatorer
    urn:nbn:se:uu:diva-70637 (URN)15070755 (PubMedID)
    Tillgänglig från: 2005-04-26 Skapad: 2005-04-26 Senast uppdaterad: 2011-01-12
    3. Multiplex amplification enabled by selective circularization of large sets of genomic DNA fragments
    Öppna denna publikation i ny flik eller fönster >>Multiplex amplification enabled by selective circularization of large sets of genomic DNA fragments
    Visa övriga...
    2005 Ingår i: Nucleic Acids Research, ISSN 1362-4962, Vol. 33, nr 8, s. e71-Artikel i tidskrift (Refereegranskat) Published
    Identifikatorer
    urn:nbn:se:uu:diva-93440 (URN)
    Tillgänglig från: 2005-09-14 Skapad: 2005-09-14Bibliografiskt granskad
    4. PieceMaker: selection of DNA fragments for selector-guided multiplex amplification
    Öppna denna publikation i ny flik eller fönster >>PieceMaker: selection of DNA fragments for selector-guided multiplex amplification
    2005 (Engelska)Ingår i: Nucleic Acids Research, ISSN 0305-1048, E-ISSN 1362-4962, Vol. 33, nr 8, s. e72-Artikel i tidskrift (Refereegranskat) Published
    Abstract [en]

    We describe PieceMaker, a software tool for the design of applications of selector probes-oligonucleotide probes that direct circularization of target nucleic acid molecules. Such probes can be combined in parallel to circularize a selection of fragments from restriction digested total genomic DNA. These fragments can then be amplified in a single PCR using a common primer pair, yielding substrates for subsequent analyses, such as parallel genotyping or sequencing. However, designing multiplex selector assays is a laborious task. The PieceMaker program alleviates this problem by selecting restriction enzymes to generate suitable fragments for selection, and generating the output data required to design the selector probes.

    Nyckelord
    Computational Biology, DNA Restriction Enzymes/metabolism, DNA; Circular/chemistry, Genomics, Oligonucleotide Probes/*chemistry, Polymerase Chain Reaction, Research Support; Non-U.S. Gov't, Software
    Nationell ämneskategori
    Medicin och hälsovetenskap
    Identifikatorer
    urn:nbn:se:uu:diva-75748 (URN)doi:10.1093/nar/gni071 (DOI)15860769 (PubMedID)
    Tillgänglig från: 2006-02-16 Skapad: 2006-02-16 Senast uppdaterad: 2017-12-14Bibliografiskt granskad
  • 8.
    Dey, Bonoshree
    et al.
    Uppsala universitet, Teknisk-naturvetenskapliga vetenskapsområdet, Biologiska sektionen, Institutionen för biologisk grundutbildning.
    Jenny, Karlsson
    Uppsala universitet, Teknisk-naturvetenskapliga vetenskapsområdet, Biologiska sektionen, Institutionen för biologisk grundutbildning.
    Yacoub, Nicole
    Uppsala universitet, Teknisk-naturvetenskapliga vetenskapsområdet, Biologiska sektionen, Institutionen för biologisk grundutbildning.
    Eklund, Marcus
    Uppsala universitet, Teknisk-naturvetenskapliga vetenskapsområdet, Biologiska sektionen, Institutionen för biologisk grundutbildning.
    Jonsson, Matilda
    Uppsala universitet, Teknisk-naturvetenskapliga vetenskapsområdet, Biologiska sektionen, Institutionen för biologisk grundutbildning.
    Alternativa Detektionsmetoder för Microarrays2016Självständigt arbete på grundnivå (kandidatexamen), 10 poäng / 15 hpStudentuppsats (Examensarbete)
  • 9.
    Garbergs, Hanna
    Uppsala universitet, Teknisk-naturvetenskapliga vetenskapsområdet, Biologiska sektionen, Institutionen för biologisk grundutbildning.
    Extraction of therapeutic proteins from dried blood spots and their analysis on Gyrolab2011Självständigt arbete på avancerad nivå (yrkesexamen), 20 poäng / 30 hpStudentuppsats (Examensarbete)
    Abstract [en]

    A method for extraction of therapeutic proteins from dried blood spots (DBS) followed by quantification on Gyrolab(TM) has been developed. The method makes it possible to measure the concentration of the analyte in the range 100-6000 ng/mL. The procedure can generate full analytical information from 15 μL blood originally sampled from a subject. The modest sample requirements allows for sampling a full pre-clinical pharmacokinetic profile from a single mouse. This may allow for reduced usage of animals during preclinical development of new therapeutic proteins in accordance with the 3R’s, replace, refine and reduce.

  • 10.
    Gustafsson, Susanne
    Uppsala universitet, Teknisk-naturvetenskapliga vetenskapsområdet, Teknisk-naturvetenskapliga fakulteten, Biologiska sektionen, Institutionen för evolutionsbiologi, Naturvårdsbiologi och genetik.
    Population genetic analyses in the orchid genus Gymnadenia: a conservation genetic perspective2003Doktorsavhandling, sammanläggning (Övrigt vetenskapligt)
    Abstract [en]

    Small populations are facing a particular risk of extinction due to a lack of appropriate genetic diversity and associated negative effects, factors dealt with in the discipline of conservation genetics. Many orchid species exhibit characteristics that make them a perfect study object in the scope of conservation genetics. The aim with this thesis was to investigate genetic structure at different levels in two orchid species Gymnadenia conopsea, geographically widespread, although diminishing and G. odoratissima with a long history of being rare. Microsatellite markers, developed in and used in studies of G. conopsea were also used in the study of G. odoratissima.

    Populations of G. conopsea expressed high levels of genetic variation and a certain amount of gene flow, although investigated mating pattern in a small population indicated non-random mating among individuals, with the majority of pollen exchange between near neighbours, and noticeable levels of geitonogamous pollinations. Further a pronounced year to year variation in flowering frequency among individuals was found.

    It was also discovered that flowering time variants (early and late) within the species G. conopsea were highly differentiated and seem to have had a more ancient historical separation than the separation between the two different species, G. conopsea and G. odoratissima.

    Levels of genetic variation in the rare congener, G. odoratissima differed between island and mainland populations where the more numerous island populations expressed larger levels of genetic variation and were less differentiated compared to the few remaining and genetically depauperated mainland populations.

    Uppsala University Library, Box 510, 75120, Uppsala, Sweden

  • 11.
    Kristoffersson, Anders
    Uppsala universitet, Teknisk-naturvetenskapliga vetenskapsområdet, Biologiska sektionen, Institutionen för biologisk grundutbildning.
    Rolling circle transcription on smallest size double stranded DNA minicircles2010Självständigt arbete på avancerad nivå (masterexamen), 20 poäng / 30 hpStudentuppsats (Examensarbete)
    Abstract [en]

    The RNA polymerase T7 is utilized as a component of motor complexes in DNA nanotechnology due to its high promotor specificity, the lack of external transcription factors and its very high processivity, but there is no experience of its application on small double stranded DNA circles. Circular templates from 210 to 126 bp in circumference sharing a common promotor termination motif were synthesized and transcription was monitored at end point on gel and in real time with a 2’ O methyl RNA molecular beacon. The RNAP T7 was found to be able to utilize circular dsDNA templates down to 126 bp with moderate impact on transcription rate for saturated systems and rolling circle transcription products were evident with denaturizing PAGE gel electrophoresis for templates down to 167 bp.

  • 12.
    Pei, Zhichao
    et al.
    Attana AB.
    Anderson, Henrik
    Uppsala universitet, Teknisk-naturvetenskapliga vetenskapsområdet, Tekniska sektionen, Institutionen för teknikvetenskaper, Fasta tillståndets elektronik.
    Myrskog, Annica
    IFM, Linköping University.
    Dunèr, Gunnar
    Attana AB.
    Ingemarsson, Björn
    Attana AB.
    Aastrup, Teodor
    Attana AB.
    Optimizing immobilization on two-dimensional carboxyl surface: pH dependence of antibody orientation and antigen binding capacity2010Ingår i: Analytical Biochemistry, ISSN 0003-2697, E-ISSN 1096-0309, Vol. 398, nr 2, s. 161-168Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    The performance of immunosensors is highly dependent on the amount of immobilized antibodies and their remaining antigen binding capacity. In this work, a method for immobilization of antibodies on a two dimensional carboxyl surface has been optimized using quartz crystal microbalance biosensors. We have shown that successful immobilization is highly dependent on surface pKa, antibody pI and pH of immobilization buffer. By use of EDC/sulfo-NHS activation reagents, the effect of the intrinsic surface pKa is avoided and immobilization also at very low pH has been made possible which is of importance for immobilization of acidic proteins. Generic immobilization conditions were demonstrated on a panel of antibodies which resulted in an average coefficient of variation of 4% for the immobilization of these antibodies.

    Antigen binding capacity as a function of immobilization pH was studied. In most cases the antigen binding capacity followed the immobilization response. However, the antigen to antibody binding ratio differed between the antibodies investigated, and for one of the antibodies, the antigen binding capacity was significantly lower than expected from immobilization in a certain pH range. Tests with anti-Fc and anti-Fab antibodies on different antibody surfaces showed that the orientation of the antibodies on the surface had a profound effect on the antigen binding capacity of the immobilized antibodies.

  • 13.
    Rundström, Gerd
    Uppsala universitet, Teknisk-naturvetenskapliga vetenskapsområdet, Kemiska sektionen, Institutionen för fysikalisk och analytisk kemi. Ytbioteknik.
    Covalent coupling of antibodies to solid phases, in particular to particles and their use in immunoassay2006Licentiatavhandling, monografi (Övrigt vetenskapligt)
    Abstract [en]

    The immobilisation of antibodies (Ab) or antigens (Ag) to solid phases is always a critical step when developing a heterogeneous immunoassay. There are two main procedures: passive adsorption and covalent coupling. Adsorption is a simple method, but it has its disadvantages including denaturation of antibodies and their leakage from the surface. The covalent coupling requires reactive group's both on the solid phase and on the protein (in this study an Ab). The most commonly used group on the protein is the ε-amino group (-NH2) on the amino acid lysine and there are many different covalent coupling techniques available for coupling to reactive groups on the solid phases. It is often advantageous to use particles as solid phase or detector reagents in heterogeneous immunoassays since this simplifies separation of bound immuno complex from free immuno reactants. Particles with a vide variety of reactive groups and diameters are suitable and easily-accessible for use as solid phase, while dyed polystyrene particles or gold particles are often used as detector reagent.

    In this work antibodies were immobilised to different solid phases, mostly particles, and the aim was to determine how the antibodies were affected by immobilisation and how this later influenced the results of an immunoassay. In paper I, Abs against Immunoglobuline E (IgE) were coupled to polystyrene particles by different covalent coupling methods and used as solid phases in an RIA determining total IgE. In the second paper (II) fluorescent polystyrene particles were used as detector reagents and antibodies against eosinophil protein X (EPX) and human neutrophil lipocaline (HNL) were coupled using two different covalent coupling techniques: The-antibodies used as capture sites in paper II were conjugated to polyethylene glycol (PEG) prior to immobilisation on a nitrocellulose membrane. The reagents were evaluated in a lateral immunochromatography (ICR) assay determining EPX and HNL. The choice of immobilisation method influences the result in the final assay and it is an advantage to have simple methods for evaluating the immobilised reagents before using them in the final assay. We thus developed different methods for evaluation of the immobilised antibodies and attempted to correlate the results from these methods with the results from the immunoassay finally used. The method used to measure the antigen binding capacity should ideally be the method of choice for evaluating differences between antibody particle derivatives. In actual fact, the results from this method did not show any subsequent correlation to the results from the immunoassay. We have therefore preferred to functionally evaluate the immobilised antibodies in the final immunoassay.

  • 14.
    Sonesson, William
    et al.
    Uppsala universitet, Teknisk-naturvetenskapliga vetenskapsområdet, Tekniska sektionen, Institutionen för teknikvetenskaper.
    Sandström Parke, Hilding
    Uppsala universitet, Teknisk-naturvetenskapliga vetenskapsområdet, Tekniska sektionen, Institutionen för teknikvetenskaper.
    The key aspects during departmental technology transfer: A case study at a biopharmaceutical company2018Självständigt arbete på avancerad nivå (masterexamen), 20 poäng / 30 hpStudentuppsats (Examensarbete)
    Abstract [en]

    In this case study the authors have tried to fill the gap of technology transferliterature focused on the biopharmaceutical industry. The technology transferliterature displays a clear industry-specific gap, mostly focused on heavy- andpharmaceutical industries. The authors have tried to find the key aspects of asuccessful technology transfer from the literature on the subject from alldifferent industries. The authors have then used these aspects to create atheoretical framework of the aspects that are possibly applicable in thebiopharmaceutical industry.

    A case study has been conducted at The Company which has a long pedigreeas one of the most innovative companies within the biopharmaceuticalindustry. The Company both develops and manufactures diagnostic tests forantibodies in animals, and their products are today widely known within theindustry. The authors have conducted a series of interviews, a non-participantobservation and also reviewed documentation of previous productsdevelopment processes. These qualitative methods have provided bothempirical evidence of similarities between the technology transfer literatureand a biopharmaceutical technology transfer process, as well as evidence ofwhat aspects are of importance in the biopharmaceutical industry. Using thisabductive research strategy, the authors have determined the key aspects thatare conceivably applicable in the biopharmaceutical industry. These are Goalcombability, Communication and documentation, Transfer plan andInterdepartmental collaboration. These aspects have not been implementedand therefore not been tested at The Company.

  • 15.
    Särnholm, Axel
    et al.
    Uppsala universitet, Teknisk-naturvetenskapliga vetenskapsområdet, Biologiska sektionen, Institutionen för biologisk grundutbildning.
    Björkesten, Johan
    Uppsala universitet, Teknisk-naturvetenskapliga vetenskapsområdet, Biologiska sektionen, Institutionen för biologisk grundutbildning.
    Zamany Company, Ayda
    Uppsala universitet, Teknisk-naturvetenskapliga vetenskapsområdet, Biologiska sektionen, Institutionen för biologisk grundutbildning.
    Berglund, Anton
    Uppsala universitet, Teknisk-naturvetenskapliga vetenskapsområdet, Biologiska sektionen, Institutionen för biologisk grundutbildning.
    Eklund, Sandra
    Uppsala universitet, Teknisk-naturvetenskapliga vetenskapsområdet, Biologiska sektionen, Institutionen för biologisk grundutbildning.
    Multiplexad Lusfältsavläsning2012Självständigt arbete på grundnivå (kandidatexamen), 10 poäng / 15 hpStudentuppsats (Examensarbete)
    Abstract [sv]

    I framtiden kommer multiplexad detektion att spela en central roll inom kliniskt bruk. Detta för att utvecklingen av riktad molekylär medicin är på uppgång och kräver studier av interaktioner mellan makromolekyler. Inom forskning används fluorescensmikroskopi för multiplexad detektion men inom kliniskt bruk föredras det att studera preparat med ljusfältsmikroskopi. Dagens problem med ljusfältsmikroskopi ligger i att det är svårt att detektera flera olika markörer samtidigt, multiplexning. Olink har lämnat en förfrågan på en ny testbar metod för multiplexad ljusfältsavläsning (bilaga 3).

    Bland dagens metoder för multiplexad analys har NuanceTM som baseras på liquid crystal tunable filter (LCTF) och SpectraLamp som baseras på agile spectral light source (ASLS) studerats och utvärderats. NuanceTM är en färdig produkt som idag finns ute på marknaden.

    Utöver att studera dagens metoder har två idéer för multiplexning formulerats av projektet. Huvudidén går under arbetsnamnen pyro immunohistochemistry (PIHC) och nyttjar teknik från pyrosekvensering för att visualisera markörer med ljusblixtar. Utöver huvudidén har projektet även ett utkast på en idé, restriction immunohistochemistry (RIHC). Den baseras på upprepad immunohistokemisk inmärkning där infärgningen avlägsnas mellan varje steg

  • 16.
    Zieba, Agata
    et al.
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för immunologi, genetik och patologi. Uppsala universitet, Science for Life Laboratory, SciLifeLab.
    Ponten, Fredrik
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för immunologi, genetik och patologi. Uppsala universitet, Science for Life Laboratory, SciLifeLab.
    Uhlen, Mathias
    KTH Royal Inst Technol, Sci Life Lab, Stockholm, Sweden..
    Landegren, Ulf
    Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för immunologi, genetik och patologi. Uppsala universitet, Science for Life Laboratory, SciLifeLab.
    In situ protein detection with enhanced specificity using DNA-conjugated antibodies and proximity ligation2018Ingår i: Modern Pathology, ISSN 0893-3952, E-ISSN 1530-0285, Vol. 31, nr 2, s. 253-263Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    Antibodies are important tools in anatomical pathology and research, but the quality of in situ protein detection by immunohistochemistry greatly depends on the choice of antibodies and the abundance of the targeted proteins. Many antibodies used in scientific research do not meet requirements for specificity and sensitivity. Accordingly, methods that improve antibody performance and produce quantitative data can greatly advance both scientific investigations and clinical diagnostics based on protein expression and in situ localization. We demonstrate here protocols for antibody labeling that allow specific protein detection in tissues via bright-field in situ proximity ligation assays, where each protein molecule must be recognized by two antibodies. We further demonstrate that single polyclonal antibodies or purified serum preparations can be used for these dual recognition assays. The requirement for protein recognition by pairs of antibody conjugates can significantly improve specificity of protein detection over single-binder assays.

1 - 16 av 16
RefereraExporteraLänk till träfflistan
Permanent länk
Referera
Referensformat
  • apa
  • ieee
  • modern-language-association
  • vancouver
  • Annat format
Fler format
Språk
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Annat språk
Fler språk
Utmatningsformat
  • html
  • text
  • asciidoc
  • rtf