TY - CHAP
T1 - Antibody Isolation from Human Synthetic Libraries of Single-Chain Antibodies and Analysis Using NGS
AU - Amir, Adi
AU - Taussig, David
AU - Bitton, Almog
AU - Nahary, Limor
AU - Vaisman-Mentesh, Anna
AU - Benhar, Itai
AU - Wine, Yariv
N1 - Publisher Copyright:
© The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature 2023.
PY - 2023
Y1 - 2023
N2 - Antibody libraries came into existence 30 years ago when the accumulating sequence data of immunoglobulin genes and the advent of PCR technology made it possible to clone antibody gene repertoires. Phage display (most common) and additional display and screening technologies were applied to pan out desired binding specificities from antibody libraries. As other antibody discovery tools, phage display is not an off-the-shelf technology and not offered as a kit but rather requires experience and expertise for making it indeed very useful. Next-generation sequencing (NGS) coupled with bioinformatics is a powerful tool for analyzing large amount of DNA sequence output of the panning. Here, we demonstrate how NGS analysis of phage biopanning (phage-Seq) of complex antibody libraries can facilitate the antibody discovery process and provide insights regarding the biopanning process (see Fig. 1).
AB - Antibody libraries came into existence 30 years ago when the accumulating sequence data of immunoglobulin genes and the advent of PCR technology made it possible to clone antibody gene repertoires. Phage display (most common) and additional display and screening technologies were applied to pan out desired binding specificities from antibody libraries. As other antibody discovery tools, phage display is not an off-the-shelf technology and not offered as a kit but rather requires experience and expertise for making it indeed very useful. Next-generation sequencing (NGS) coupled with bioinformatics is a powerful tool for analyzing large amount of DNA sequence output of the panning. Here, we demonstrate how NGS analysis of phage biopanning (phage-Seq) of complex antibody libraries can facilitate the antibody discovery process and provide insights regarding the biopanning process (see Fig. 1).
KW - CDRs (complementarity determining regions)
KW - FR (variable framework region)
KW - HTS (high-throughput sequencing)
KW - NGS (next-generation sequencing
KW - Phage display
KW - Phage-Seq
KW - Single-chain antibodies
KW - Synthetic library
KW - VH (variable region of antibody heavy chain)
KW - VL (variable region of antibody light chain)
KW - scFv (single-chain variable fragment)
UR - http://www.scopus.com/inward/record.url?scp=85170179652&partnerID=8YFLogxK
U2 - 10.1007/978-1-0716-3381-6_18
DO - 10.1007/978-1-0716-3381-6_18
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C2 - 37679629
AN - SCOPUS:85170179652
T3 - Methods in Molecular Biology
SP - 347
EP - 372
BT - Methods in Molecular Biology
PB - Humana Press Inc.
ER -