2001 Fiscal Year Final Research Report Summary
Cloning and production of human anti-B monoclonal antibody in ABO blood type
| Project/Area Number |
10670403
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Legal medicine
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| Research Institution | Yamagata University (1999-2001)
Tokai University (1998) |
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Principal Investigator |
OGAWA Motoki Yamagata University School of Medicine, Department of Forensic Medicine, Professor, 医学部, 教授 (90213686)
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| Co-Investigator(Kenkyū-buntansha) |
TAKAHASHI Hiroshi Yamagata University School of Medicine, Department of Forensic Medicine, Instructor, 医学部, 助手 (10142217)
TAKEICHI Sanae Tokai University School of Medicine, Department of Forensic Medicine, Professor, 医学部, 教授 (20035497)
YUKAWA Nobuhiro Tokai University School of Medicine, Department of Forensic Medicine, Associate Professor, 医学部, 助教授 (30240154)
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| Project Period (FY) |
1998 – 2000
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| Keywords | ABO histoblood group / natural antibody / human monoclonal antibody / ELISA / flow cytmetry |
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| Research Abstract |
In our work, cloning of the variable region of human antibodies directed to B-antigen in ABO blood group, and production of the recombinant Fab fragment in Escherichia coli cells were carried out. After the culture of peripheral lymphocyte transformed by Epstein-Barr virus, specific antibody-producing cells were screened by ELISA and flowcytometry. After reverse transcription-PCR using subclass-specific primers, cDNA library was constructed and screened with a plasmid vector, pFab-His2, in the E. coli expression system. Two clones specifically reacted with B-antigen in ELISA and hemagglutination. Their nucleotide sequence of the heavy and light chains of the anti-B human antibodies was determined to be V_L1 and V_H4, respectively, in the immunoglobulin subgroup. The recombinant antibodies to the blood group antigen structure may be adaptable for phenotyping, which will provide a valuable means for forensic applications.
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