Streamlined human antibody generation and optimization by exploiting designed immunoglobulin loci in a B cell line
Open Access
- 1 June 2021
- journal article
- research article
- Published by Springer Science and Business Media LLC in Cellular & Molecular Immunology
- Vol. 18 (6), 1545-1561
- https://doi.org/10.1038/s41423-020-0440-9
Abstract
Monoclonal antibodies (mAbs) are widely utilized as therapeutic drugs for various diseases, such as cancer, autoimmune diseases, and infectious diseases. Using the avian-derived B cell line DT40, we previously developed an antibody display technology, namely, the ADLib system, which rapidly generates antigen-specific mAbs. Here, we report the development of a human version of the ADLib system and showcase the streamlined generation and optimization of functional human mAbs. Tailored libraries were first constructed by replacing endogenous immunoglobulin genes with designed human counterparts. From these libraries, clones producing full-length human IgGs against distinct antigens can be isolated, as exemplified by the selection of antagonistic mAbs. Taking advantage of avian biology, effective affinity maturation was achieved in a straightforward manner by seamless diversification of the parental clones into secondary libraries followed by single-cell sorting, quickly affording mAbs with improved affinities and functionalities. Collectively, we demonstrate that the human ADLib system could serve as an integrative platform with unique diversity for rapid de novo generation and optimization of therapeutic or diagnostic antibody leads. Furthermore, our results suggest that libraries can be constructed by introducing exogenous genes into DT40 cells, indicating that the ADLib system has the potential to be applied for the rapid and effective directed evolution and optimization of proteins in various fields beyond biomedicine.Funding Information
- New Energy and Industrial Technology Development Organization
This publication has 51 references indexed in Scilit:
- Harnessing Gene Conversion in Chicken B Cells to Create a Human Antibody Sequence RepertoirePLOS ONE, 2013
- Coupling mammalian cell surface display with somatic hypermutation for the discovery and maturation of human antibodiesProceedings of the National Academy of Sciences of the United States of America, 2011
- Development of a novel mammalian cell surface antibody display platformmAbs, 2010
- An ex vivo method for rapid generation of monoclonal antibodies (ADLib system)Nature Protocols, 2006
- Activation-Induced Cytidine Deaminase Initiates Immunoglobulin Gene Conversion and Hypermutation by a Common IntermediatePLoS Biology, 2004
- Production of Human Monoclonal and Polyclonal Antibodies in TransChromo AnimalsCloning and Stem Cells, 2002
- Fully synthetic human combinatorial antibody libraries (HuCAL) based on modular consensus frameworks and CDRs randomized with trinucleotidesJournal of Molecular Biology, 2000
- Biodiamonds and paying back debtsNature Biotechnology, 1996
- Guiding the Selection of Human Antibodies from Phage Display Repertoires to a Single Epitope of an AntigenNature Biotechnology, 1994
- Replacing the complementarity-determining regions in a human antibody with those from a mouseNature, 1986