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Construction of synthetic antibody phage-display libraries
KTH, School of Biotechnology (BIO), Protein Technology. University of Toronto, Canada.ORCID iD: 0000-0002-6104-6446
2018 (English)In: Phage Display, Humana Press, 2018, p. 45-60Chapter in book (Refereed)
Abstract [en]

Synthetic antibody libraries provide a vast resource of renewable antibody reagents that can rival or exceed those of natural antibodies and can be rapidly isolated through controlled in vitro selections. Use of highly optimized human frameworks enables the incorporation of defined diversity at positions that are most likely to contribute to antigen recognition. This protocol describes the construction of synthetic antibody libraries based on a single engineered human autonomous variable heavy domain scaffold with diversity in all three complementarity-determining regions. The resulting libraries can be used to generate recombinant domain antibodies for a wide range of protein antigens using phage display. Furthermore, analogous methods can be used to construct antibody libraries based on larger antibody fragments or second-generation libraries aimed to fine-tune antibody characteristics including affinity, specificity, and manufacturability. The procedures rely on standard reagents and equipment available in most molecular biology laboratories.

Place, publisher, year, edition, pages
Humana Press, 2018. p. 45-60
Series
Methods in Molecular Biology, ISSN 1064-3745 ; 1701
Keywords [en]
Antibody fragment, Degenerate oligonucleotide, Domain antibody, Human antibody, Phage display, Protein engineering
National Category
Biochemistry and Molecular Biology
Identifiers
URN: urn:nbn:se:kth:diva-218302DOI: 10.1007/978-1-4939-7447-4_3Scopus ID: 2-s2.0-85033695515ISBN: 978-1-4939-7446-7 (print)OAI: oai:DiVA.org:kth-218302DiVA, id: diva2:1160435
Funder
Swedish Research Council
Note

QC 20171127

Available from: 2017-11-27 Created: 2017-11-27 Last updated: 2017-11-27Bibliographically approved

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Nilvebrant, Johan

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  • nn-NB
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