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Generation and diversification of recombinant monoclonal antibodies

DeLuca, Keith F, Mick, Jeanne E, Ide, Amy H, Lima, Wanessa C, Sherman, Lori, Schaller, Kristin L, Anderson, Steven M, Zhao, Ning +4 more·2021·eLife
DOI 10.7554/elife.72093PMC PMC8763395

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Abstract

Antibodies are indispensable tools used for a large number of applications in both foundational and translational bioscience research; however, there are drawbacks to using traditional antibodies generated in animals. These include a lack of standardization leading to problems with reproducibility, high costs of antibodies purchased from commercial sources, and ethical concerns regarding the large number of animals used to generate antibodies. To address these issues, we have developed practical methodologies and tools for generating low-cost, high-yield preparations of recombinant monoclonal antibodies and antibody fragments directed to protein epitopes from primary sequences. We describe these methods here, as well as approaches to diversify monoclonal antibodies, including customization of antibody species specificity, generation of genetically encoded small antibody fragments, and conversion of single chain antibody fragments (e.g. scFv) into full-length, bivalent antibodies. This study focuses on antibodies directed to epitopes important for mitosis and kinetochore function; however, the methods and reagents described here are applicable to antibodies and antibody fragments for use in any field.

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Authors
DeLuca, K. F., Mick, J. E., Ide, A. H., Lima, W. C., Sherman, L., Schaller, K. L., Anderson, S. M., Zhao, N., Stasevich, T. J., Varma, D., Nilsson, J., & DeLuca, J. G.
Affiliations
  • Department of Biochemistry and Molecular Biology, Colorado State University
  • Geneva Antibody Facility, Faculty of Medicine, University of Geneva
  • CU Cancer Center Cell Technologies Shared Resource, University of Colorado Cancer Center, Anschutz Medical Campus
  • Department of Pediatric Hematology, Oncology and Bone Marrow Transplant, University of Colorado Anschutz Medical Campus
  • Department of Cell and Developmental Biology, Feinberg School of Medicine, Northwestern University
  • The Novo Nordisk Foundation Center for Protein Research, University of Copenhagen, Faculty of Health and Medical Sciences
ORCID
Venue
eLife
Publisher
eLife Sciences Publications, Ltd
Published
2021-12-31
Volume
10
ISSN
2050-084X
DOI
10.7554/elife.72093
PMC
PMC8763395
Cited by
21
References
2
Language
en
Licence
Funders
  • National Institute of General Medical Sciences (R35GM130365, MIRA R35GM119728, K99GM141453, R01GM135391)
  • National Science Foundation (MCB-1845761)
  • National Cancer Institute (P30CA046934)
  • Novo Nordisk Foundation Center for Protein Research (NNF14CC0001)

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