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Cited by 307Monoclonal and Polyclonal Antibodies ResearchChemical Synthesis and Analysisvaccines and immunoinformatics approaches

In vitro selection and affinity maturation of antibodies from a naive combinatorial immunoglobulin library.

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Gram, H, Marconi, L A, Barbas, C F, Collet, T A, Lerner, R A, Kang, A S·1992·Proceedings of the National Academy of Sciences
DOI 10.1073/pnas.89.8.3576PMC PMC48911
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Abstract

We have used a combinatorial immunoglobulin library approach to obtain monoclonal antibodies from nonimmune adult mice, thereby establishing the principles of (i) accessing naive combinatorial antibody libraries for predetermined specificities and (ii) increasing the affinity of the selected antibody binding sites by random mutagenesis. A combinatorial Fab library expressing immunoglobulin mu and kappa light-chain fragments on the surface of filamentous phage was prepared from bone marrow of nonimmunized, adult BALB/c mice with the multivalent display vector pComb8. Phage displaying low affinity Fabs (binding constants, 10(4)-10(5) M-1) binding to a progesterone-bovine serum albumin conjugate were isolated from the library. Random mutagenesis of the heavy- and light-chain variable regions expressed in the mono-valent phage display vector pComb3 was performed by error-prone PCR, and subsequently clones with improved affinity for the hapten conjugate were selected. We demonstrate that antibodies with desirable characteristics from a nonimmune source may be selected and affinity maturation may be achieved by using the twin vectors pComb8 and pComb3, thus opening the route to obtaining specific antibodies from a generic library and bypassing immunization.

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  • In vitro affinity maturation methods, such as random mutagenesis with display technologies, improve antibody binding against target proteins but are time consuming and labor intensive.

    “In vitro affinity maturation methods, such as random mutagenesis with display technologies, has been shown to improve antibody binding against target proteins, but such approaches are time consuming and labor intensive (27–32).”

    From Deep learning guided optimization of human antibody against SARS-CoV-2 variants with broad neutralization (2022) · cites this paper as [27]

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Research works printed but not linked: 10

  1. [6]Orlandi, R., Gussow, D. H., Jones, P. T. & Winter, G. (1989) Proc. Natl. Acad. Sci. USA 86, 3833–3837.
  2. [8]Sastry, L., Alting-Mees, M., Huse, W. D., Short, J. M., Sorge, J. A., Hay, B. N., Janda, K. D., Benkovic, S. J. & Lerner, R. A. (1989) Proc. Natl. Acad. Sci. USA 86, 5728–5732.
  3. [11]Caton, A. J. & Koprowski, H. (1990) Proc. Natl. Acad. Sci. USA 87, 6450–6454.
  4. [12]Persson, M. A., Cao, R. H. & Burton, D. R. (1991) Proc. Natl. Acad. Sci. USA 88, 2432–2436.
  5. [15]Kang, A. S., Barbas, C. F., Janda, K. D., Benkovic, S. J. & Lerner, R. A. (1991) Proc. Natl. Acad. Sci. USA 88, 4363– 4366.
  6. [17]Kang, A. S., Jones, T. M. & Burton, D. R. (1991) Proc. Natl. Acad. Sci. USA 88, 11120–11123.
  7. [16]Barbas, C. F., Kang, A. S., Lerner, R. A. & Benkovic, S. J. (1991) Proc. Natl. Acad. Sci. USA 88, 7978–7982.
  8. [18]Burton, D. R., Barbas, C. F., Persson, M. A., Koenig, S., Chanock, R. M. & Lerner, R. A. (1991) Proc. Natl. Acad. Sci. USA 88, 10134–10137.
  9. [40]Steele, E. J., Pollard, J. W., Taylor, L. & Both, G. W. (1991) in Somatic Hypermutation in V-Regions, ed. Steele, E. J. (CRC, Boca Raton, FL), pp. 137–149.
  10. [41]Diamond, B. & Scharff, M. D. (1984) Proc. Natl. Acad. Sci. USA 81, 5841–5844.

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Authors
Gram, H., Marconi, L. A., Barbas, C. F., Collet, T. A., Lerner, R. A., & Kang, A. S.
Affiliations
Scripps Research Institute (US)
ORCID
Venue
Proceedings of the National Academy of Sciences
Publisher
National Academy of Sciences
Published
1992-04-15
Volume
89
Issue
8
Pages
3576-3580
ISSN
  • 0027-8424 (ISSN-L)
  • 1091-6490
DOI
10.1073/pnas.89.8.3576
PMC
PMC48911
OpenAlex
W1969943007
Cited by
307
References
37
Citations by year
2025: 3 · 2024: 6 · 2023: 6 · 2022: 5 · 2021: 4 · 2020: 1 · 2019: 6 · 2018: 8 · 2017: 4 · 2016: 6 · 2015: 9 · 2014: 7 · 2013: 6 · 2012: 11
FWCI
28.31
Type
journal article
Language
en
Open access
Locations
  • Proceedings of the National Academy of Sciences (publishedVersion)
  • Europe PMC (PubMed Central) (publishedVersion)
  • PubMed Central (publishedVersion)
  • PubMed (publishedVersion)
Topics
  • Health Sciences › Medicine › Radiology, Nuclear Medicine and Imaging › Monoclonal and Polyclonal Antibodies Research
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Chemical Synthesis and Analysis
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › vaccines and immunoinformatics approaches
Keywords
affinity maturation, naive antibody library, phage display, Fab fragment, error-prone PCR mutagenesis, monoclonal antibody selection, variable region, bone marrow
MeSH terms
Amino Acid Sequence, Animals, Antibodies, Monoclonal, Base Sequence, Bone Marrow, Enzyme-Linked Immunosorbent Assay, Genes, Immunoglobulin, Immunoglobulin Variable Region, Immunoglobulin Fab Fragments, Immunoglobulin kappa-Chains, Immunoglobulin mu-Chains, Male, Mice, Inbred BALB C, Molecular Sequence Data, Oligodeoxyribonucleotides, Plasmids, Sequence Homology, Nucleic Acid, Restriction Mapping, Gene Library, Polymerase Chain Reaction, Mutagenesis, Mice

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