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Application of artificial gel antibodies for investigating molecular polymorphisms of human pituitary growth hormone
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Pharmaceutical Biosciences. (Fred Nyberg)
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Pharmaceutical Biosciences.
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Pharmaceutical Biosciences. (Fred Nyberg)
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Pharmaceutical Biosciences.
2011 (English)In: Amino Acids, ISSN 0939-4451, E-ISSN 1438-2199, Vol. 40, no 4, 1249-1255 p.Article in journal (Refereed) Published
Abstract [en]

Artificial gel-antibodies were used to probe human growth hormone (GH) activity in hormone preparations purified from human pituitaries. A partially purified fraction containing differently sized forms of the hormone was further processed to yield monomeric and dimeric forms of GH activity suitable for synthesizing artificial polyacrylamide gel antibodies. These two types of GH antibodies were used for probing GH activity in experiments analyzing the two forms of the hormone by HPLC gel-permeation and ion-exchange chromatography. In the size-exclusion experiments both hormone fractions eluted as homogenous peaks, whereas the ion exchanger resolved the hormones in several active components. The antibodies towards the GH monomer were more potent to recognize monomeric GH compared to antibodies against dimeric GH. The opposite was found for the dimeric GH antibodies. It was concluded that these two sets of antibodies it might be useful for discriminating between dimeric and monomeric GH in samples of clinical origin.

Place, publisher, year, edition, pages
2011. Vol. 40, no 4, 1249-1255 p.
Keyword [en]
Assay, Electrophoresis, Growth hormone (GH), HPLC, Human, Pituitary, Polyacrylamide gel antibodies, Purification, Quantification
National Category
Pharmaceutical Sciences
Research subject
Pharmaceutical Biochemistry
Identifiers
URN: urn:nbn:se:uu:diva-122448DOI: 10.1007/s00726-011-0840-3ISI: 000288546700021PubMedID: 21312046OAI: oai:DiVA.org:uu-122448DiVA: diva2:310166
Available from: 2010-04-16 Created: 2010-04-13 Last updated: 2017-12-12Bibliographically approved
In thesis
1. Application of Artificial Gel Antibodies for the Detection and Quantification of Proteins in Biological Fluids
Open this publication in new window or tab >>Application of Artificial Gel Antibodies for the Detection and Quantification of Proteins in Biological Fluids
2010 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

The molecular-imprinting method has previously been used for the synthesis of artificial gel antibodies, highly selective for various proteins. In present study, we have synthesized artificial gel antibodies against haemoglobin, albumin and different forms of growth hormone with the aim to develop a simple and rapid procedure to measure the concentration of these protein biomarkers in samples of clinical interest.  A spectrophotometric method was developed to design a standard curve in the form of a straight line, whereby the true absorption (not the recorded “apparent” absorption) was plotted against a known protein concentration. The procedure, applied to quantitative analysis of albumin in human plasma and cerebrospinal fluid (CSF) from patients with ALS, indicated that  the concentration of this protein was significantly enhanced in CSF from patients with amyotrophic lateral sclerosis (ALS), compared to control samples. A low level of albumin was observed in plasma from ALS patients compared to controls. Additionally, free zone electrophoresis was employed to detect human growth hormone (GH) activity in hormone preparations purified from human pituitaries. We have successfully synthesized antibodies capable of discriminating between dimeric and monomeric GH in samples of clinical origin. To quantify these proteins a calibration curve has been designed, i.e. a plot of the electrophoretic mobility of the complex GH/gel antibody against the protein concentration in the sample, for instance serum or CSF.

This method was also employed for qualitative and quantitative determinations of Somatropin, a non-glycosylated GH and glycosylated-GH in a body liquid.

Our results indicate that by this technique one can “fish out” with high accuracy various proteins from both body fluids containing a great number of other proteins. It might well apply also to biomarker proteins for other diseases.

Place, publisher, year, edition, pages
Uppsala: Acta Universitatis Upsaliensis, 2010. 64 p.
Series
Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Science and Technology, ISSN 1651-6214 ; 741
National Category
Medicinal Chemistry Pharmaceutical Sciences
Research subject
Pharmaceutical Science
Identifiers
urn:nbn:se:uu:diva-122457 (URN)978-91-554-7802-5 (ISBN)
Public defence
2010-05-18, C4:305, Uppsala biomedicinska centrum BMC, Husarg. 3, Uppsala, 13:00 (English)
Opponent
Supervisors
Available from: 2010-04-27 Created: 2010-04-13 Last updated: 2010-05-17Bibliographically approved

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Ghasemzadeh, Nasim

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