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Screening of a human antibody phage display library against a peptide antigen using stimuli-responsive bioconjugates

Title data

Stocker, Gisela ; Dumoulin, Dorothée ; Vandevyver, Caroline ; Hilbrig, Frank ; Freitag, Ruth:
Screening of a human antibody phage display library against a peptide antigen using stimuli-responsive bioconjugates.
In: Biotechnology Progress. Vol. 24 (November 2008) Issue 6 . - pp. 1314-1324.
ISSN 1520-6033
DOI: https://doi.org/10.1002/btpr.60

Official URL: Volltext

Abstract in another language

A synthetic antibody phage library (ETH-2) was screened against the MUC-1 peptide. Two standard methods were used, namely screening on immunotubes coated with the peptide antigen and on streptavidin-coated paramagnetic particles together with a biotinylated MUC-1 peptide. The results were compared with those obtained using a novel approach based on a stimuli-responsive bioconjugate of avidin and poly-(N-isopropylacrylamide), also in combination with the biotinylated peptide. The poly-(N-isopropylacrylamide)-based bioconjugate is soluble below a temperature of 30 degrees C in the aqueous solutions pertinent to this study, but precipitates rapidly once this temperature is surpassed. The process is reversible, and the bioconjugate redissolves once the temperature is lowered again. The stimuli-responsive bioconjugate was used to isolate specific phage from the library in several rounds of panning, consisting of repeated thermoprecipitation/redissolution cycles in a procedure known as "affinity precipitation." As far as we know, this is the first time a compound as big as a viral particle has been specifically captured by a stimuli-responsive bioconjugate. Compared to the two other investigated screening techniques, affinity precipitation of the phage led to a greater variety of genetically different specific phage, while the process was easy to handle and required no dedicated equipment safe for a centrifuge and a thermostated water bath.

Further data

Item Type: Article in a journal
Refereed: Yes
Keywords: affinity precipitation; lower critical solution temperature; polyNIPAAm; phage display; stimuli-responsive materials
Institutions of the University: Faculties > Faculty of Engineering Science
Faculties > Faculty of Engineering Science > Chair Process Biotechnology
Faculties > Faculty of Engineering Science > Chair Process Biotechnology > Chair Process Biotechnology - Univ.-Prof. Dr. Ruth Freitag
Faculties
Result of work at the UBT: No
DDC Subjects: 600 Technology, medicine, applied sciences
600 Technology, medicine, applied sciences > 620 Engineering
Date Deposited: 24 Feb 2016 13:36
Last Modified: 24 Feb 2016 13:36
URI: https://eref.uni-bayreuth.de/id/eprint/31035