Class-switching of the IgM type anti-adenocarcinoma human antibody HB4C5 into an IgG1 type resulted in the loss of the antigen binding ability

Hirotaka Haruta, Hirofumi Tachibana, Katsumi Mochizuki, Shuichi Hashizume, Kiyoko Kusakabe, Sanetaka Shirahata, Hiroki Murakami

Research output: Contribution to journalArticle

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Abstract

The heavy and light chain genes (μ and λ) of the human anti-adenocarcinoma monoclonal antibody HB4C5 (MAb-C5) were cloned from the human-human hybridoma cell line HB4C5 and co-expressed in Chinese hamster ovary (CHO) and COS-7 cells. ELISA assay and the RI imaging of the cancer tissue xenografted into nude mice showed that the recombinant MAb-C5 retained the original antigen binding activity. We then replaced the IgM constant region of this MAb-C5 heavy chain gene with the human IgG1 constant region gene and co-expressed it with the light chain gene. This recombination was confirmed by a complete DNA sequencing and Western-blot analysis. Despite the fact that the DNA sequence, the expressed protein size, and the assembly of heavy and light chains were indicated to be normal, the IgG1 type MAb-C5 could not bind to the original antigen. This result suggests that this antibody alters its antigen binding property upon class-switching.

Original languageEnglish
Pages (from-to)137-145
Number of pages9
JournalHuman Antibodies
Volume8
Issue number3
Publication statusPublished - 1997

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Immunoglobulin Class Switching
Adenocarcinoma
Immunoglobulin G
Monoclonal Antibodies
Antigens
Antibodies
Light
Genes
COS Cells
Hybridomas
Cricetulus
DNA Sequence Analysis
Nude Mice
Genetic Recombination
Immunoglobulin M
Ovary
Western Blotting
Enzyme-Linked Immunosorbent Assay
Cell Line
anti-IgM

All Science Journal Classification (ASJC) codes

  • Immunology and Allergy
  • Immunology

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Class-switching of the IgM type anti-adenocarcinoma human antibody HB4C5 into an IgG1 type resulted in the loss of the antigen binding ability. / Haruta, Hirotaka; Tachibana, Hirofumi; Mochizuki, Katsumi; Hashizume, Shuichi; Kusakabe, Kiyoko; Shirahata, Sanetaka; Murakami, Hiroki.

In: Human Antibodies, Vol. 8, No. 3, 1997, p. 137-145.

Research output: Contribution to journalArticle

Haruta, H, Tachibana, H, Mochizuki, K, Hashizume, S, Kusakabe, K, Shirahata, S & Murakami, H 1997, 'Class-switching of the IgM type anti-adenocarcinoma human antibody HB4C5 into an IgG1 type resulted in the loss of the antigen binding ability', Human Antibodies, vol. 8, no. 3, pp. 137-145.
Haruta, Hirotaka ; Tachibana, Hirofumi ; Mochizuki, Katsumi ; Hashizume, Shuichi ; Kusakabe, Kiyoko ; Shirahata, Sanetaka ; Murakami, Hiroki. / Class-switching of the IgM type anti-adenocarcinoma human antibody HB4C5 into an IgG1 type resulted in the loss of the antigen binding ability. In: Human Antibodies. 1997 ; Vol. 8, No. 3. pp. 137-145.
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T1 - Class-switching of the IgM type anti-adenocarcinoma human antibody HB4C5 into an IgG1 type resulted in the loss of the antigen binding ability

AU - Haruta, Hirotaka

AU - Tachibana, Hirofumi

AU - Mochizuki, Katsumi

AU - Hashizume, Shuichi

AU - Kusakabe, Kiyoko

AU - Shirahata, Sanetaka

AU - Murakami, Hiroki

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AB - The heavy and light chain genes (μ and λ) of the human anti-adenocarcinoma monoclonal antibody HB4C5 (MAb-C5) were cloned from the human-human hybridoma cell line HB4C5 and co-expressed in Chinese hamster ovary (CHO) and COS-7 cells. ELISA assay and the RI imaging of the cancer tissue xenografted into nude mice showed that the recombinant MAb-C5 retained the original antigen binding activity. We then replaced the IgM constant region of this MAb-C5 heavy chain gene with the human IgG1 constant region gene and co-expressed it with the light chain gene. This recombination was confirmed by a complete DNA sequencing and Western-blot analysis. Despite the fact that the DNA sequence, the expressed protein size, and the assembly of heavy and light chains were indicated to be normal, the IgG1 type MAb-C5 could not bind to the original antigen. This result suggests that this antibody alters its antigen binding property upon class-switching.

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