In Vitro Antibody Affinity Maturation Targeting Germline Hotspots
Open accessAbstract
Affinity-matured antibodies can exhibit increased biological efficacy. Regardless of whether an antibody is isolated from a hybridoma or a human Fv phage library, the antibody affinity for its target may need improvement for therapeutic applications. An increased affinity may allow for a reduced dosage of a therapeutic antibody; toxic side effects may also be reduced. In the immune system, affinity maturation is a process involving somatic hypermutations in B cells. Therefore, germline hotspot residues are most likely to have a major impact on antibody affinity. Here, we describe procedures for germline hotspot mutagenesis with an emphasis on strategies for randomizing hotspots with PCR and phage display, using as an example the anti-CD22 monoclonal antibody.
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Integrity signals
Registry
Cites
Statistics
Claims the paper makes
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Claims other papers make about this one
Each is checked against this paper’s own text: does it say what the citing paper says it does?
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 [30]
Self-check
Percentages, sums and reported differences re-computed from the paper’s own numbers.
The self-check needs the paper’s full text.
Cites
Continuous cultures of fused cells secreting antibody of predefined specificity
Köhler, G., & Milstein, C. (1975) · Nature · doi:10.1038/256495a0 · Cited by 17,200 · see citing works
Sequences of proteins of immunological interest
Kabat, B. E. A. (1984) · Analytical Biochemistry · doi:10.1016/0003-2697(84)90805-4 · Cited by 3,363 · see citing works
Rapid generation of functional human IgG antibodies derived from Fab-on-phage display libraries
Jostock, T., Vanhove, M., Brepoels, E., van Gool, R., Daukandt, M., Wehnert, A., van Hegelsom, R., Dransfield, D., Sexton, D., Devlin, M., et al. (2004) · Journal of Immunological Methods · doi:10.1016/j.jim.2004.03.014 · Cited by 2,562 · see citing works
Assembly of combinatorial antibody libraries on phage surfaces: the gene III site.
Barbas, C. F., Kang, A. S., Lerner, R. A., & Benkovic, S. J. (1991) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.88.18.7978 · Cited by 1,183 · see citing works
Hoogenboom, H. R., Griffiths, A. D., Johnson, K. S., Chiswell, D. J., Hudson, P. J., & Winter, G. P. (1991) · Nucleic Acids Research · doi:10.1093/nar/19.15.4133 · Cited by 1,019 · see citing works
Phage Display : A Laboratory Manual
Barbas, C. F. (2001) · Medical Entomology and Zoology · Cited by 952 · see citing works
IMGT, the international ImMunoGeneTics database
Ruiz, M. (2000) · Nucleic Acids Research · doi:10.1093/nar/28.1.219 · Cited by 859 · see citing works
Kinetic and affinity limits on antibodies produced during immune responses.
Foote, J., & Eisen, H. N. (1995) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.92.5.1254 · Cited by 375 · see citing works
Display technologies: Application for the discovery of drug and gene delivery agents☆
Sergeeva, A., Kolonin, M. G., Molldrem, J. J., Pasqualini, R., & Arap, W. (2006) · Advanced Drug Delivery Reviews · doi:10.1016/j.addr.2006.09.018 · Cited by 266 · see citing works
Neuberger, M. S., & Milstein, C. (1995) · Current Opinion in Immunology · doi:10.1016/0952-7915(95)80010-7 · Cited by 259 · see citing works
Improving antibody affinity by mimicking somatic hypermutation in vitro
Chowdhury, P. S., & Pastan, I. H. (1999) · Nature Biotechnology · doi:10.1038/9872 · Cited by 256 · see citing works
Self-citationEngineering of therapeutic antibodies to minimize immunogenicity and optimize function
Presta, L. G. (2006) · Advanced Drug Delivery Reviews · doi:10.1016/j.addr.2006.01.026 · Cited by 247 · see citing works
IMGT, the international ImMunoGeneTics database
Lefranc, M. (2001) · Nucleic Acids Research · doi:10.1093/nar/29.1.207 · Cited by 221 · see citing works
WAM: an improved algorithm for modelling antibodies on the WEB
Whitelegg, N., & Rees, A. R. (2000) · Protein Engineering Design and Selection · doi:10.1093/protein/13.12.819 · Cited by 159 · see citing works
Salvatore, G., Beers, R., Margulies, I., Kreitman, R. J., & Pastan, I. H. (2002) · PubMed · Cited by 153 · see citing works
Self-citationMonitoring and interpreting the intrinsic features of somatic hypermutation
Neuberger, M. S., Ehrenstein, M. R., Klix, N., Jolly, C. J., Yélamos, J., Rada, C. A., & Milstein, C. (1998) · Immunological Reviews · doi:10.1111/j.1600-065x.1998.tb01434.x · Cited by 118 · see citing works
Ho, M., Kreitman, R. J., Onda, M., & Pastan, I. H. (2004) · Journal of Biological Chemistry · doi:10.1074/jbc.m409783200 · Cited by 114 · see citing works
Self-citationGenome-wide somatic hypermutation
Wang, C. L., Harper, R. A., & Wabl, M. R. (2004) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.0402009101 · Cited by 111 · see citing works
The targeting of somatic hypermutation
Jolly, C. J., Wagner, S. D., Rada, C. A., Klix, N., Milstein, C., & Neuberger, M. S. (1996) · Seminars in Immunology · doi:10.1006/smim.1996.0020 · Cited by 101 · see citing works
Beers, R., Chowdhury, P., Bigner, D. D., & Pastan, I. (2000) · PubMed · Cited by 75 · see citing works
Self-citationLigand-assisted Aggregation of Proteins
Ho, J., Kitov, P., Paszkiewicz, E., Sadowska, J., Bundle, D., & Ng, K. (2005) · Journal of Biological Chemistry · doi:10.1074/jbc.m504403200 · Cited by 29 · see citing works
Flagged first, then by citations
Cited by
Sorted by citations
Shan, S., Luo, S., Yang, Z., Hong, J., Su, Y., Ding, F., Fu, L., Li, C., Chen, P., Ma, J., et al. (2022) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.2122954119 · Cited by 157 · see citing works
Li, N., Fu, H., Hewitt, S. M., Dimitrov, D. S., & Ho, M. (2017) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.1706055114 · Cited by 132 · see citing works
Self-citationThe role of phage display in therapeutic antibody discovery
Chan, C. E. Z., Lim, A. P. C., MacAry, P. A., & Hanson, B. J. (2014) · International Immunology · doi:10.1093/intimm/dxu082 · Cited by 87 · see citing works
Feng, M., Bian, H.-J., Wu, X., Fu, T., Fu, Y., Hong, J., Fleming, B. D., Flajnik, M. F., & Ho, M. (2018) · Antibody Therapeutics · doi:10.1093/abt/tby011 · Cited by 82 · see citing works
Self-citationA novel high‐affinity human monoclonal antibody to mesothelin
Ho, M., Feng, M., Fisher, R. J., Rader, C. M., & Pastan, I. H. (2010) · International Journal of Cancer · doi:10.1002/ijc.25557 · Cited by 60 · see citing works
Self-citationTang, Z., Feng, M., Gao, W., Phung, Y., Chen, W., Chaudhary, A., Croix, B. S., Qian, M., Dimitrov, D. S., & Ho, M. (2013) · Molecular Cancer Therapeutics · doi:10.1158/1535-7163.mct-12-0731 · Cited by 48 · see citing works
Self-citationLim, C. C., Choong, Y. S., & Lim, T. S. (2019) · International Journal of Molecular Sciences · doi:10.3390/ijms20081861 · Cited by 44 · see citing works
Generation of recombinant antibodies and means for increasing their affinity
Altshuler, E. P., Serebryanaya, D. V., & Katrukha, A. (2010) · Biochemistry (Moscow) · doi:10.1134/s0006297910130067 · Cited by 32 · see citing works
Xu, M., Lei, G., Chen, M., Wang, K., Lv, W., Zhang, P., Hu, T., Gao, J., Lü, C., Mei, Y., et al. (2020) · EBioMedicine · doi:10.1016/j.ebiom.2021.103250 · Cited by 21 · see citing works
Gorman, J., Cheung, C. S., Duan, Z., Ou, L., Wang, M., Chen, X., Cheng, C., Biju, A., Sun, Y., Wang, P., et al. (2024) · Nature Communications · doi:10.1038/s41467-023-44534-y · Cited by 20 · see citing works
Self-citationLi, T., Shen, Y., Lin, F., Fu, W., Liu, S., Wang, C., Liang, J., Fan, X., Ye, X., Tang, Y., et al. (2019) · The FASEB Journal · doi:10.1096/fj.201802354r · Cited by 12 · see citing works
Kraivong, R., Traewachiwiphak, S., Nilchan, N., Tangthawornchaikul, N., Pornmun, N., Poraha, R., Sriruksa, K., Limpitikul, W., Avirutnan, P., Malasit, P., et al. (2022) · PLoS ONE · doi:10.1371/journal.pone.0266136 · Cited by 10 · see citing works
Yang, Q. Z., Wu, G., Jin, J., Wang, H., Zhang, J., Liu, L., Zhao, H., Wang, J., & Yang, X. (2023) · BMC Biotechnology · doi:10.1186/s12896-023-00776-8 · Cited by 9 · see citing works
A General Sequence Processing and Analysis Program for Protein Engineering
Stafford, R., Zimmerman, E. S., Hallam, T. J., & Sato, A. K. (2014) · Journal of Chemical Information and Modeling · doi:10.1021/ci500362s · Cited by 9 · see citing works
Generative Diffusion Models for Antibody Design, Docking, and OptimizationPreprint
Peng, Z., Han, C., Wang, X., Li, D., & Yuan, F. (2023) · bioRxiv (Cold Spring Harbor Laboratory) · doi:10.1101/2023.09.25.559190 · Cited by 9 · see citing works
Immunochemical engineering of cell surfaces to generate virus resistance
Xie, J., Sok, D., Wu, N. C., Zheng, T., Zhang, W., Burton, D. R., & Lerner, R. A. (2017) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.1702764114 · Cited by 5 · see citing works
Qudsia, S., Merugu, S. B., Mangukiya, H. B., Negi, H., Wu, Z., & Li, D. (2018) · Biochemical and Biophysical Research Communications · doi:10.1016/j.bbrc.2018.03.131 · Cited by 5 · see citing works
Nur, A., Lai, J. Y., Ch’ng, A. C. W., Choong, Y. S., Isa, W. Y. H. W., & Lim, T. S. (2024) · International Journal of Biological Macromolecules · doi:10.1016/j.ijbiomac.2024.134217 · Cited by 2 · see citing works
Germline Genetics in Cancer: The New Frontier
Milanese, J., & Wang, E. (2020) · Elsevier eBooks · doi:10.1016/b978-0-12-801238-3.11667-8 · Cited by 1 · see citing works
High Affinity Maturated Human Antibodies from Naïve and Synthetic Antibody Repertoires
Lim, C. C., Choong, Y. S., & Lim, T. S. (2018) · InTech eBooks · doi:10.5772/intechopen.71664 · Cited by 1 · see citing works
Evolution of Bioaffinity Reagents by Phage Display
Brockmann, E.-C. (2010) · UTUPub (University of Turku)
- Authors
- Ho, M., & Pastan, I.
- Affiliations
- National Institutes of Health (US)
- Corresponding author
- Ho, Mitchell
- Venue
- Methods in Molecular Biology™
- Publisher
- Humana Press
- Published
- 2008-12-10
- Volume
- 525
- Pages
- 293-308
- ISSN
- 1064-3745 (ISSN-L)
- 1940-6029
- PMC
- PMC3473074
- OpenAlex
- W10761404
- Cited by
- 21
- References
- 21
- Citations by year
- 2024: 2 · 2023: 2 · 2022: 2 · 2020: 2 · 2019: 2 · 2018: 3 · 2017: 2 · 2014: 2 · 2013: 1
- FWCI
- 4.83
- Type
- book chapter
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Locations
- Methods in molecular biology (publishedVersion)
- PubMed (publishedVersion)
- PubMed Central (acceptedVersion)
- Funders
- National Institutes of Health (Z99 CA999999)
- National Cancer Institute (Z01BC010891, Y99 CA999999, ZIABC010891)
- Topics
- Health Sciences › Medicine › Radiology, Nuclear Medicine and Imaging › Monoclonal and Polyclonal Antibodies Research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein purification and stability
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Chemical Synthesis and Analysis
- Keywords
- affinity maturation, somatic hypermutation, anti-CD22 monoclonal antibody, phage display, random mutagenesis, therapeutic antibodies, B cells
- MeSH terms
- Amino Acid Sequence, Antibodies, Monoclonal, Antibody Affinity, Bacteriophages, Base Sequence, Enzyme-Linked Immunosorbent Assay, Escherichia coli, Flow Cytometry, Germ Cells, Models, Molecular, Molecular Biology, Molecular Sequence Data, Sequence Alignment, Sequence Analysis, DNA, Electroporation, Peptide Library
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
Are you an author, or do you see a problem with this page? Request a review