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Phage display screening in low dielectric media
University of Kalmar, School of Pure and Applied Natural Sciences.
University of Kalmar, School of Pure and Applied Natural Sciences.
University of Kalmar, School of Pure and Applied Natural Sciences.
University of Kalmar, School of Pure and Applied Natural Sciences.ORCID iD: 0000-0002-0407-6542
2008 (English)In: Journal of Molecular Recognition, ISSN 0952-3499, E-ISSN 1099-1352, Vol. 21, no 5, p. 330-337Article in journal (Refereed) Published
Abstract [en]

Here we report the first application of phage display screening in low dielectric media. Two series of phage clones with affinity for α-chymotrypsin (CT) were selected from a Ph.D.TM-C7C library, using either a buffer or acetonitrile in buffer (50%, v/v). The affinity of lysates, individual clones or selected cyclic peptides for the enzyme was studied by examining their influence on CT activity. Peptides displayed on phage selected in buffer provided significant protection from enzyme autolysis resulting in marked increase in CT activity (>100%). Phage selected in ACN provided some, albeit weak, protection from the detrimental influence on CT from ACN. In conclusion, the results demonstrate the potential for the application of phage display screening protocols to targets in media of low dielectricity.

Place, publisher, year, edition, pages
2008. Vol. 21, no 5, p. 330-337
National Category
Organic Chemistry
Research subject
Natural Science, Organic Chemistry; Natural Science, Biotechnology
Identifiers
URN: urn:nbn:se:lnu:diva-1808DOI: 10.1002/jmr.904OAI: oai:DiVA.org:lnu-1808DiVA, id: diva2:308856
Available from: 2010-04-06 Created: 2010-04-06 Last updated: 2017-12-12Bibliographically approved
In thesis
1. Towards novel applications for biomolecular interactions at surfaces
Open this publication in new window or tab >>Towards novel applications for biomolecular interactions at surfaces
2013 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

The focus of this thesis has been to examine two new concepts involving the interaction of biological agents with synthetic material surfaces.

The feasibility of deploying phage display techniques for the screening of materials in non-biological media was examined in two studies. Firstly, the utility of a phage library based on the M13 phage was used to identify peptides with affinity for α-chymotrypsin in media containing significant amounts of an organic solvent, acetonitrile. The selected peptides were demonstrated to inhibit enzyme autolysis. The influence of immobilization to the substrate used in the screening studies, Eupergit®, on the function of this enzyme in organic solvents was examined; enhanced enzyme resilience to organic solvents was demonstrated. Secondly, phage display screening for cyclic heptapeptide motifs selective for adenine was undertaken using an adenine derivatized glass surfaces as a target. A peptide with affinity for adenine was identified and quartz crystal microbalance studies demonstrated selectivity for the adeninyl moiety.

Finally, the influence of oseltamivir molecularly imprinted polymer particles on the capacity of avian influenza A virus H1N1 to replicate in Madin-Darby Canine Kidney cells was investigated. The polymer matrix was shown to demonstrate a general though small suppression of virus replication.

Place, publisher, year, edition, pages
Växjö: Linnaeus University Press, 2013
Series
Linnaeus University Dissertations ; 149/2013
Keywords
molecular recognition, interactions, phage display, biomolecular interactions, phage, adenine, influenza virus replication, oseltamivir, imprinted polymers.
National Category
Biomaterials Science
Identifiers
urn:nbn:se:lnu:diva-29505 (URN)978-91-87427-53-4 (ISBN)
Public defence
2013-10-25, N2007, Västergård, Smålandsgatan 26E, Kalmar, 09:30 (English)
Opponent
Available from: 2013-10-08 Created: 2013-10-08 Last updated: 2016-11-24Bibliographically approved

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Olofsson, LinusSöderberg, PernillaAnkarloo, JonasNicholls, Ian Alan

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