Streamlined human antibody generation and optimization by exploiting designed immunoglobulin loci in a B cell line

Autor: Kunihiro Ohta, Shu-ichi Hashimoto, Chika Koyama, Ke-Yi Lin, Hitomi Masuda, Yukoh Nakazaki, Atsushi Sawada, Kenjiro Asagoshi, Tomoaki Uchiki, Kanako Tamai, Kohei Kurosawa, Shigehisa Kawata, Takashi Yabuki, Goh Sasaki, Aki Takaiwa, Shunsuke Miyai, Naoki Takahashi, Hidetaka Seo
Rok vydání: 2020
Předmět:
Vascular Endothelial Growth Factor A
0301 basic medicine
Cell biology
medicine.drug_class
Immunology
Gene Conversion
Gene Dosage
homologous recombination
Computational biology
Hydroxamic Acids
Monoclonal antibody
Article
Antibodies
drug discovery
Cell Line
Affinity maturation
03 medical and health sciences
0302 clinical medicine
Antigen
medicine
Animals
Humans
Immunology and Allergy
Amino Acid Sequence
Gene
antibody therapeutics
Immunoglobulin rearrangements
B-Lymphocytes
antibody engineering
Base Sequence
biology
Tumor Necrosis Factor-alpha
Drug discovery
Antibodies
Monoclonal
Genetic Variation
Directed evolution
Antibodies
Neutralizing
030104 developmental biology
Infectious Diseases
030220 oncology & carcinogenesis
Antibody Formation
biology.protein
Antibody
Homologous recombination
Chickens
Pseudogenes
Zdroj: Cellular and Molecular Immunology
ISSN: 2042-0226
1672-7681
Popis: Monoclonal antibodies (mAbs) are widely utilized as therapeutic drugs for various diseases, such as cancer, autoimmune diseases, and infectious diseases. Using the avian-derived B cell line DT40, we previously developed an antibody display technology, namely, the ADLib system, which rapidly generates antigen-specific mAbs. Here, we report the development of a human version of the ADLib system and showcase the streamlined generation and optimization of functional human mAbs. Tailored libraries were first constructed by replacing endogenous immunoglobulin genes with designed human counterparts. From these libraries, clones producing full-length human IgGs against distinct antigens can be isolated, as exemplified by the selection of antagonistic mAbs. Taking advantage of avian biology, effective affinity maturation was achieved in a straightforward manner by seamless diversification of the parental clones into secondary libraries followed by single-cell sorting, quickly affording mAbs with improved affinities and functionalities. Collectively, we demonstrate that the human ADLib system could serve as an integrative platform with unique diversity for rapid de novo generation and optimization of therapeutic or diagnostic antibody leads. Furthermore, our results suggest that libraries can be constructed by introducing exogenous genes into DT40 cells, indicating that the ADLib system has the potential to be applied for the rapid and effective directed evolution and optimization of proteins in various fields beyond biomedicine.
Databáze: OpenAIRE
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