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A unique method for antibody to possess the catalytic function (3 rd report)
Author(s) -
Uda Taizo,
Akiyoshi Yuko,
Taguchi Hiroaki,
Hifumi Emi
Publication year - 2018
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2018.32.1_supplement.lb77
Subject(s) - monoclonal antibody , antibody , antigen , peptide , immunoglobulin light chain , chemistry , catalysis , biochemistry , substrate (aquarium) , microbiology and biotechnology , peptide sequence , gene , biology , genetics , ecology
Since a natural catalytic antibody found at 1989, many mouse‐ and human‐type of catalytic antibodies cleaving the targeting antigens such as peptides, nucleotides, viruses and bacterial proteins, molecules such as factor VIII, CCR5, TNFα etc. have been produced so far (1, 2). In contrast, thousands of monoclonal antibodies (mAbs) have been produced over past 40 years. However, not all monoclonal antibodies have catalytic activity. It is considered that the proportion possessing a catalytic function is about several percentages or lower. Thus, the authors have continued to explore the technology how to put the catalytic function to those normal (non‐catalytic) mAbs. Recently, we have made a significant progress in part with respect to the technology. After the genes of human antibody light chain were inserted into pET20b (+), E. coli was transformed and each gene was expressed by inducing with IPTG. After recovering the soluble fraction of the culture supernatant, it was purified by Ni‐NTA‐, cation‐, and/or size exclusion‐chromatography. The enzyme activity was measured using Arg(R)‐pNA as a synthetic substrate and/or the antigens such as amyloid‐beta (Aβ) peptide etc. Since We found interesting two light chains, #7TR and #7GY. The former could degrade both FRET‐Aβ substrate and Aβ1–40 full peptide. In contrast, #7GY, whose sequence is identical to that of #7TR except for the amino acids of the positions at 29 th and 30 th , did not degrade the FRET‐Aβ substrate and Aβ1–40 full peptide at all. The chemical features of the two light chains were investigated in detail by using a synthetic substrate, Arg‐pNA. #7TR showed higher hydrolytic activity for Arg‐pNA substrate than that of #7GY by a factor of one and half. In addition, the presence of Zn (II) ion hugely enhanced the catalytic activity of #7TR light chain but not #7GY. Fluorescence spectroscopy clearly exhibited the difference of the two antibody light chains. Through these facts and discussion, we will propose a clue how to we can put the catalytic function to a normal (non‐catalytic) antibody. This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .