uu.seUppsala University Publications
Change search
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • ieee
  • modern-language-association
  • vancouver
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf
Protein-Nanocellulose Interactions in Paper Filters for Advanced Separation Applications
Uppsala University, Disciplinary Domain of Science and Technology, Technology, Department of Engineering Sciences, Nanotechnology and Functional Materials.
Uppsala University, Disciplinary Domain of Science and Technology, Technology, Department of Engineering Sciences, Nanotechnology and Functional Materials.
Uppsala University, Disciplinary Domain of Science and Technology, Technology, Department of Engineering Sciences, Nanotechnology and Functional Materials.ORCID iD: 0000-0002-8105-2317
2017 (English)In: Langmuir, ISSN 0743-7463, E-ISSN 1520-5827, Vol. 33, no 19, p. 4729-4736Article in journal (Refereed) Published
Abstract [en]

Protein-based pharmaceutics are widely explored for healthcare applications, and 6 out of 10 best-selling drugs today are biologicals. The goal of this work was to evaluate the protein nanocellulose interactions in paper filter for advanced separation applications such as virus removal filtration and bioprocessing. The protein recovery was measured for bovine serum albumin (BSA), gamma-globulin, and lysozyme using biuret total protein reagent and polyacrylamide gel electrophoresis (PAGE), and the throughput was characterized in terms of flux values from fixed volume filtrations at various protein concentrations and under worst case experimental conditions. The affinity of cellulose to bind various proteins, such as BSA, lysozyme, gamma-globulin, and human IgG was quantified using a quartz crystal microbalance (QCMB) by developing a new method of fixing the cellulose fibers to the electrode surface without cellulose dissolution-precipitation. It was shown that the. mille-feuille filter exhibits high protein recovery, that is, similar to 99% for both BSA and lysozyme. However, gamma-globulin does not pass through the membrane due to its large size (i.e., >180 kDa). The PAGE data show no substantial change in the amount of dimers and trimers before and after filtration. QCMB analysis suggests a low affinity between the nanocellulose surface and proteins. The nanocellulose-based filter exhibits desirable inertness as a filtering material intended for protein purification.

Place, publisher, year, edition, pages
AMER CHEMICAL SOC , 2017. Vol. 33, no 19, p. 4729-4736
National Category
Nano Technology
Research subject
Engineering Science with specialization in Nanotechnology and Functional Materials
Identifiers
URN: urn:nbn:se:uu:diva-326244DOI: 10.1021/acs.langmuir.7b00566ISI: 000401674900015PubMedID: 28441870OAI: oai:DiVA.org:uu-326244DiVA, id: diva2:1130586
Funder
Knut and Alice Wallenberg FoundationStiftelsen Olle Engkvist ByggmästareAvailable from: 2017-08-10 Created: 2017-08-10 Last updated: 2018-10-24
In thesis
1. Mille-feuille Filter: A Non-woven Nano-cellulose Based Virus Removal Filter for Bioprocessing
Open this publication in new window or tab >>Mille-feuille Filter: A Non-woven Nano-cellulose Based Virus Removal Filter for Bioprocessing
2018 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Virus removal filters, produced from synthetic surface-modified polymers or regenerated cellulose by phase inversion, are vital to the production of therapeutic proteins such as monoclonal antibodies and plasma proteins. Use of these filters is also one of the most expensive purification steps in the downstream processing of proteins due to high sales price and being limited to a single use.

In this thesis, a virus removal filter produced from Cladophora sp. algal nanocellulose has been characterized. The mille-feuille (‘‘a thousand leaves’’) filter paper is the first non-woven, wet-laid filter paper composed of 100% native nanocellulose that is capable of removing the ‘‘worst-case’’ model viruses, the non-enveloped parvoviruses, i.e., minute virus of mice (MVM; 18–20 nm), from water with a log10 reduction value (LRV) ≥5.78 (≥99.9998%). The mille-feuille filter features a unique internal stratified architecture that is the result of nanofiber self-assembly into 2D nanosheets during manufacturing. Such an internal structure has several benefits for achieving highly selective virus removal with high flux.

The pore size distribution can be tailored to sizes from 10 to 25 nm by altering drying conditions, i.e. temperature and drying rate; therefore, the filter can be customized to target the size cut-off of the smallest virus particles known. The mille-feuille filter has achieved up to 200 L m-2 h-1 (LMH) bar-1 in flux. Furthermore, protein recovery rates of 99% were measured during bovine serum albumin (BSA) filtration. Protein recovery was determined to be dependent on the protein size and charge.

Filtration of cell culture media was also investigated, and no fouling was observed with fluxes of 400 LMH for an 11 µm filter and 140 LMH for a 33 µm filter at 3 bar. An LRV of >4.8 was measured for the 33 µm filter at 3 bar, but only 2.2 was measured for the 11 µm filter at 3 bar using the small-size ФX174 bacteriophage as a model virus.

Furthermore, the virus reduction was discovered to be pressure dependent, with the LRV increasing with trans membrane pressure (TMP). The tendency to virus breakthrough was partly mitigated at low TMPs by filter cross-linking.

In summary, the mille-feuille filter paper has the characteristics to be a promising virus removal filter for both upstream and downstream applications. Further studies shall focus on the area of protein filtration to gain a better understanding of how buffer conditions and the physical characteristics of proteins contribute to filter fouling.

Place, publisher, year, edition, pages
Uppsala: Acta Universitatis Upsaliensis, 2018. p. 70
Series
Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Science and Technology, ISSN 1651-6214 ; 1738
Keywords
virus filtration, mille-feuille, downstream, nanocellulose, protein throughput, protein recovery, LRV, MVM, Cladophora, non-woven filter
National Category
Nano Technology
Identifiers
urn:nbn:se:uu:diva-364082 (URN)978-91-513-0489-2 (ISBN)
Public defence
2018-12-14, Polhemsalen, 10134, Ångström, Lägerhyddsvägen 1, Uppsala, 09:15 (English)
Opponent
Supervisors
Available from: 2018-11-20 Created: 2018-10-24 Last updated: 2018-11-30

Open Access in DiVA

No full text in DiVA

Other links

Publisher's full textPubMed

Authority records BETA

Gustafsson, SimonMihranyan, Albert

Search in DiVA

By author/editor
Gustafsson, SimonManukyan, LevonMihranyan, Albert
By organisation
Nanotechnology and Functional Materials
In the same journal
Langmuir
Nano Technology

Search outside of DiVA

GoogleGoogle Scholar

doi
pubmed
urn-nbn

Altmetric score

doi
pubmed
urn-nbn
Total: 258 hits
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • ieee
  • modern-language-association
  • vancouver
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf