Next Article in Journal
Use of Monoclonal Antibody to Treat COVID-19 in Children and Adolescents: Risk of Abuse of Prescription and Exacerbation of Health Inequalities
Next Article in Special Issue
Research and Clinical Landscape of Bispecific Antibodies for the Treatment of Solid Malignancies
Previous Article in Journal
G-Quadruplex-Based Drug Delivery Systems for Cancer Therapy
Previous Article in Special Issue
Antibody–Drug Conjugate to Treat Meningiomas
 
 
Search for Articles:
Title / Keyword
Author / Affiliation / Email
Journal
Article Type
 
 
Advanced Search
 
Section
Special Issue
Volume
Issue
Number
Page
 
Journals
Pharmaceuticals
Volume 14
Issue 7
10.3390/ph14070672
pharmaceuticals-logo
Submit to this Journal Review for this Journal Propose a Special Issue
► Article Menu

Article Menu

share Share announcement Help format_quote Cite question_answer Discuss in SciProfiles
first_page
settings
Order Article Reprints
Font Type:
Arial Georgia Verdana
Font Size:
Aa Aa Aa
Line Spacing:
Column Width:
Background:
Article

Site-Specific Antibody Conjugation to Engineered Double Cysteine Residues

by
Qun Zhou
1,*,
Josephine Kyazike
1,
Ekaterina Boudanova
1,
Michael Drzyzga
1,
Denise Honey
1,
Robert Cost
1,
Lihui Hou
1,
Francis Duffieux
2,
Marie-Priscille Brun
3,
Anna Park
1 and
Huawei Qiu
1
1
Large Molecules Research, Sanofi, Framingham, MA 01701, USA
2
Large Molecules Research, Sanofi, 94400 Vitry-Sur-Seine, France
3
Integrated Drug Discovery, Sanofi, 94400 Vitry-Sur-Seine, France
*
Author to whom correspondence should be addressed.
Pharmaceuticals 2021, 14(7), 672; https://doi.org/10.3390/ph14070672
Submission received: 2 June 2021 / Revised: 7 July 2021 / Accepted: 9 July 2021 / Published: 14 July 2021
(This article belongs to the Special Issue Evaluation of the Antitumor Mechanism of Armed Antibodies)
Browse Figures
Review Reports Versions Notes

Abstract

Site-specific antibody conjugations generate homogeneous antibody-drug conjugates with high therapeutic index. However, there are limited examples for producing the site-specific conjugates with a drug-to-antibody ratio (DAR) greater than two, especially using engineered cysteines. Based on available Fc structures, we designed and introduced free cysteine residues into various antibody CH2 and CH3 regions to explore and expand this technology. The mutants were generated using site-directed mutagenesis with good yield and properties. Conjugation efficiency and selectivity were screened using PEGylation. The top single cysteine mutants were then selected and combined as double cysteine mutants for expression and further investigation. Thirty-six out of thirty-eight double cysteine mutants display comparable expression with low aggregation similar to the wild-type antibody. PEGylation screening identified seventeen double cysteine mutants with good conjugatability and high selectivity. PEGylation was demonstrated to be a valuable and efficient approach for quickly screening mutants for high selectivity as well as conjugation efficiency. Our work demonstrated the feasibility of generating antibody conjugates with a DAR greater than 3.4 and high site-selectivity using THIOMABTM method. The top single or double cysteine mutants identified can potentially be applied to site-specific antibody conjugation of cytotoxin or other therapeutic agents as a next generation conjugation strategy.
Keywords: site-specific antibody-drug conjugation; THIOMABTM; engineered double cysteine; PEGylation; conjugation efficiency and selectivity site-specific antibody-drug conjugation; THIOMABTM; engineered double cysteine; PEGylation; conjugation efficiency and selectivity

Share and Cite

MDPI and ACS Style

Zhou, Q.; Kyazike, J.; Boudanova, E.; Drzyzga, M.; Honey, D.; Cost, R.; Hou, L.; Duffieux, F.; Brun, M.-P.; Park, A.; et al. Site-Specific Antibody Conjugation to Engineered Double Cysteine Residues. Pharmaceuticals 2021, 14, 672. https://doi.org/10.3390/ph14070672

AMA Style

Zhou Q, Kyazike J, Boudanova E, Drzyzga M, Honey D, Cost R, Hou L, Duffieux F, Brun M-P, Park A, et al. Site-Specific Antibody Conjugation to Engineered Double Cysteine Residues. Pharmaceuticals. 2021; 14(7):672. https://doi.org/10.3390/ph14070672

Chicago/Turabian Style

Zhou, Qun, Josephine Kyazike, Ekaterina Boudanova, Michael Drzyzga, Denise Honey, Robert Cost, Lihui Hou, Francis Duffieux, Marie-Priscille Brun, Anna Park, and et al. 2021. "Site-Specific Antibody Conjugation to Engineered Double Cysteine Residues" Pharmaceuticals 14, no. 7: 672. https://doi.org/10.3390/ph14070672

APA Style

Zhou, Q., Kyazike, J., Boudanova, E., Drzyzga, M., Honey, D., Cost, R., Hou, L., Duffieux, F., Brun, M.-P., Park, A., & Qiu, H. (2021). Site-Specific Antibody Conjugation to Engineered Double Cysteine Residues. Pharmaceuticals, 14(7), 672. https://doi.org/10.3390/ph14070672

Note that from the first issue of 2016, this journal uses article numbers instead of page numbers. See further details here.

Article Metrics

Back to TopTop