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  Magnetic bead-based semi-automated phage display panning strategy for the directed evolution of antibodies

Ch'ng, A. C. W., Konthur, Z., & Lim, T. S. (2020). Magnetic bead-based semi-automated phage display panning strategy for the directed evolution of antibodies. Methods in Enzymology, 630, 159-178. doi:10.1016/bs.mie.2019.10.023.

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Item Permalink: http://hdl.handle.net/21.11116/0000-0005-9AAB-D Version Permalink: http://hdl.handle.net/21.11116/0000-0006-86F3-0
Genre: Journal Article

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 Creators:
Ch'ng, Angela Chiew Wen, Author
Konthur, Zoltán1, Author              
Lim, Theam Soon, Author
Affiliations:
1Zoltán Konthur, Biomolekulare Systeme, Max Planck Institute of Colloids and Interfaces, Max Planck Society, ou_1863499              

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Free keywords: Directed evolution, Phage display, Magnetic nanoparticles, Antibody, Semi-automated biopanning, KingFisher Flex, Phage library
 Abstract: Directed evolution is a proven approach to fine tune or modify biomolecules for various applications ranging from research to industry. The process of evolution requires methods that are capable of not only generating genetic diversity but also to distinguish the variants of desired characteristics. One method that is synonymous with directed evolution of proteins is phage display. Here, we present a protocol describing the application of magnetic nanoparticles coupled with a processor to carry out the identification of monoclonal antibodies (mAbs) from a diverse antibody library via phage display. Target antigens are coupled to magnetic nanoparticles as the solid phase for the isolation of the binding mAbs via affinity. A gradual enrichment in clones would result in increasing ELISA readouts with increasing rounds of panning. During monoclonal level analysis, positivity can be deduced with comparison to background and controls. The biopanning process can also be adopted for the directed evolution of enzymes, scaffold proteins or even peptides.

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Language(s): eng - English
 Dates: 2019-10-312020
 Publication Status: Published in print
 Pages: -
 Publishing info: -
 Table of Contents: -
 Rev. Method: -
 Identifiers: DOI: 10.1016/bs.mie.2019.10.023
Other: Artikel angefragt 5.2.20 SN
BibTex Citekey: CHNG2020159
 Degree: -

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Title: Methods in Enzymology
Source Genre: Journal
 Creator(s):
Affiliations:
Publ. Info: New York : Academic Press
Pages: - Volume / Issue: 630 Sequence Number: - Start / End Page: 159 - 178 Identifier: ISSN: 0076-6879

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Title: Nanoarmoring of Enzymes with Carbon Nanotubes and Magnetic Nanoparticles
Source Genre: Book
 Creator(s):
Kumar, Challa V., Editor
Affiliations:
-
Publ. Info: -
Pages: - Volume / Issue: - Sequence Number: - Start / End Page: - Identifier: ISBN: 978-0-12-820143-5