Improvement of Biophysical Properties and Affinity of a Human Anti-L1CAM Therapeutic Antibody through Antibody Engineering Based on Computational Methods

Autor: Heesu Chae, Seonjoo Yoon, Seulki Cho, Jisu Hong, Jiwoo Lee, Jae Young Lee, Munsik Jeong, Hyojeong Hong, Dongwook Choi, Woosuk Nam, Kiyoung Kwon
Rok vydání: 2021
Předmět:
0301 basic medicine
Models
Molecular
L1
Chemical Phenomena
Antibody Affinity
Drug Evaluation
Preclinical
Protein Engineering
030226 pharmacology & pharmacy
biophysical properties
computational methods
0302 clinical medicine
Biology (General)
Spectroscopy
biology
antibody engineering
Chemistry
Protein Stability
Immunogenicity
Antibodies
Monoclonal
General Medicine
Computer Science Applications
Biochemistry
therapeutic antibody
Therapeutic antibody
Thermodynamics
medicine.symptom
Antibody
Cell bank
medicine.drug_class
QH301-705.5
Neural Cell Adhesion Molecule L1
CHO Cells
Monoclonal antibody
Catalysis
Article
Inorganic Chemistry
03 medical and health sciences
Cricetulus
In vivo
anti-cancer antibody
medicine
Animals
Humans
research cell bank
Physical and Theoretical Chemistry
Molecular Biology
QD1-999
Organic Chemistry
030104 developmental biology
Mechanism of action
Drug Design
biology.protein
Zdroj: International Journal of Molecular Sciences
International Journal of Molecular Sciences, Vol 22, Iss 6696, p 6696 (2021)
Volume 22
Issue 13
ISSN: 1422-0067
Popis: The biophysical properties of therapeutic antibodies influence their manufacturability, efficacy, and safety. To develop an anti-cancer antibody, we previously generated a human monoclonal antibody (Ab417) that specifically binds to L1 cell adhesion molecule with a high affinity, and we validated its anti-tumor activity and mechanism of action in human cholangiocarcinoma xenograft models. In the present study, we aimed to improve the biophysical properties of Ab417. We designed 20 variants of Ab417 with reduced aggregation propensity, less potential post-translational modification (PTM) motifs, and the lowest predicted immunogenicity using computational methods. Next, we constructed these variants to analyze their expression levels and antigen-binding activities. One variant (Ab612)—which contains six substitutions for reduced surface hydrophobicity, removal of PTM, and change to the germline residue—exhibited an increased expression level and antigen-binding activity compared to Ab417. In further studies, compared to Ab417, Ab612 showed improved biophysical properties, including reduced aggregation propensity, increased stability, higher purification yield, lower pI, higher affinity, and greater in vivo anti-tumor efficacy. Additionally, we generated a highly productive and stable research cell bank (RCB) and scaled up the production process to 50 L, yielding 6.6 g/L of Ab612. The RCB will be used for preclinical development of Ab612.
Databáze: OpenAIRE
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