Chemically crosslinked bispecific antibodies for cancer therapy: Breaking from the structural restrictions of the genetic fusion approach

Asami Ueda, Mitsuo Umetsu, Takeshi Nakanishi, Kentaro Hashikami, Hikaru Nakazawa, Shuhei Hattori, Ryutaro Asano, Izumi Kumagai

Research output: Contribution to journalArticle

Abstract

Antibodies are composed of structurally and functionally independent domains that can be used as building blocks to construct different types of chimeric protein-format molecules. However, the generally used genetic fusion and chemical approaches restrict the types of structures that can be formed and do not give an ideal degree of homogeneity. In this study, we combined mutation techniques with chemical conjugation to construct a variety of homogeneous bivalent and bispecific antibodies. First, building modules without lysine residues—which can be chemical conjugation sites—were generated by means of genetic mutation. Specific mutated residues in the lysine-free modules were then re-mutated to lysine residues. Chemical conjugation at the recovered lysine sites enabled the construction of homogeneous bivalent and bispecific antibodies from block modules that could not have been so arranged by genetic fusion approaches. Molecular evolution and bioinformatics techniques assisted in finding viable alternatives to the lysine residues that did not deactivate the block modules. Multiple candidates for re-mutation positions offer a wide variety of possible steric arrangements of block modules, and appropriate linkages between block modules can generate highly bioactive bispecific antibodies. Here, we propose the effectiveness of the lysine-free block module design for site-specific chemical conjugation to form a variety of types of homogeneous chimeric protein-format molecule with a finely tuned structure and function.

Original languageEnglish
Article number711
JournalInternational journal of molecular sciences
Volume21
Issue number3
DOIs
Publication statusPublished - 2020 Feb 1

Keywords

  • Antibody
  • Bioinformatics
  • Chemical conjugation

ASJC Scopus subject areas

  • Catalysis
  • Molecular Biology
  • Spectroscopy
  • Computer Science Applications
  • Physical and Theoretical Chemistry
  • Organic Chemistry
  • Inorganic Chemistry

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