A novel method of preparing the monoform structure of catalytic antibody light chain

Emi Hifumi, Shingo Matsumoto, Hiroki Nakashima, Shogo Itonaga, Mitsue Arakawa, Yoshiki Katayama, Ryuichi Kato, Taizo Uda

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Abstract

Along with the development of antibody drugs and catalytic antibodies, the structural diversity (heterogeneity) of antibodies has been given attention. For <20 yr, detailed studies on the subject have not been conducted, because the phenomenon presents many difficult and complex problems. Structural diversity provides some (or many) isoforms of an antibody distinguished by different charges, different molecular sizes, and modificationsof amino acid residues. For practicaluse, the antibody and the subunits must have a defined structure. In recent work, we have found that the copper (Cu) ion plays a substantial role in solving the diversity problem. In the current study, we used several catalytic antibody light chains to examine the effect of the Cu ion. In all cases, the different electrical charges of the molecule converged to a single charge, giving 1 peak in cation-exchange chromatography, as well as a single spot in 2-dimensional gel electrophoresis. The Cu-binding site was investigated by using mutagenesis, ultraviolet-visible spectroscopy, atomic force microscope analysis, and molecular modeling, which suggested that histidine and cysteine residues close to the C-terminus are involved with the binding site. The constant region domain of the antibody light chain played an important role in the heterogeneity of the light chain. Our findings may be a significant tool for preparing a single defined, not multiple, isoform structure.-Hifumi, E., Matsumoto, S., Nakashima, H., Itonaga, S., Arakawa, M., Katayama, Y., Kato, R., Uda T. A novel method of preparing the monoform structure of catalytic antibody light chain.

Original languageEnglish
Pages (from-to)895-908
Number of pages14
JournalFASEB Journal
Volume30
Issue number2
DOIs
Publication statusPublished - Feb 1 2016

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All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Biochemistry
  • Molecular Biology
  • Genetics

Cite this

Hifumi, E., Matsumoto, S., Nakashima, H., Itonaga, S., Arakawa, M., Katayama, Y., ... Uda, T. (2016). A novel method of preparing the monoform structure of catalytic antibody light chain. FASEB Journal, 30(2), 895-908. https://doi.org/10.1096/fj.15-276394