Rational library design by functional CDR resampling

•A synthetic antibody fragment library resampling the functional CDRs was constructed.•The resultant library yielded de novo specific hits against a wide spectrum of targets. Successful antibody discovery relies on diversified libraries, where two aspects are implied, namely the absolute number of u...

Full description

Saved in:
Bibliographic Details
Published in:New biotechnology 2018-10, Vol.45, p.89-97
Main Authors: Zhao, Qi, Buhr, Diane, Gunter, Courtney, Frenette, Jenny, Ferguson, Mary, Sanford, Eric, Holland, Erika, Rajagopal, Chitra, Batonick, Melissa, Kiss, Margaret M., Weiner, Michael P.
Format: Article
Language:English
Subjects:
Antibodies
Antibodies - genetics
Antibodies - isolation & purification
Antibodies - metabolism
Biocompatibility
Bioinformatics
Combinatorial analysis
Complementarity Determining Regions - genetics
Complementarity Determining Regions - isolation & purification
Complementarity Determining Regions - metabolism
Display devices
Humans
Immunoglobulins
Immunology
Libraries
Peptide Library
Phage display
Phages
Proteins
Resampling
Technology
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:•A synthetic antibody fragment library resampling the functional CDRs was constructed.•The resultant library yielded de novo specific hits against a wide spectrum of targets. Successful antibody discovery relies on diversified libraries, where two aspects are implied, namely the absolute number of unique clones and the percentage of functional clones. Instead of pursuing the absolute quantity thresholded by current display technology, we have sought to maximize the effective diversity by improving functional clone percentage. With the combined effort of bioinformatics, structural biology, molecular immunology and phage display technology, we devised a bioinformatic pipeline to construct and validate libraries via combinatorial assembly of sequences from a database of experimentally validated antibodies. Furthermore, we showed that the libraries constructed as such yielded a significantly increased success rate against different antigen types and generated over 20-fold more unique hits per targets compared with libraries based on traditional degenerate nucleotide methods. Our study indicated that predefined CDR sequences with optimized CDR-framework compatibility could be a productive direction of functional library construction for in vitro antibody development.
ISSN:1871-6784
1876-4347
DOI:10.1016/j.nbt.2017.12.005