Grant-in-Aid for international Scientific Research
|Allocation Type||Single-year Grants|
|Research Institution||Osaka University|
TOMITA Ken-ichi Professor, faculty of Pharmaceutical Sciences, Osaka University., 薬学部, 教授 (30028831)
AKANMORI Bar カ゛ーナ大学, 野口英世記念医学研究所(カ゛ーナ、レゴン), 研究員
ARMAH George E. Research Fellow, Noguchi Memorial Institute for Medical Research, University of, 野口英世記念医学研究所(カ゛ーナ、レゴン), 研究員
POLJAK Roberto J. Professor, Institut Pasteur, 25 rue du Dr. Roux, Paris Cedex 15, France., 免疫構造部門(フランス、ハ゜リー), 教授
HORII Toshihiro Associate Professor, research Institute for Microbial Diseases, Osaka University, 微生物病研究所, 助教授 (80142305)
OMATA Yoshitaka Research Associate, Department of Veterinary Physiology and Protozoan Immunology, 原虫病分子免疫研究センター, 助手 (10132987)
UESUGI Seiichi Associate Professor, Faculty of Pharmaceutical Sciences, Osaka University. (Pres, 工学部, 教授 (70028851)
YAMAGATA Yuriko Research Associate, Faculty of Pharmaceutical Sciences, Osaka University., 薬学部, 助手 (40183678)
HAKOSHIMA. Toshio Research Associate, Faculty of Pharmaceutical Sciences, Osaka University., 薬学部, 助手 (00164773)
FUJII Satoshi Associate Professor, faculty of Pharmaceutical sciences, Osaka University., 薬学部, 助教授 (10107104)
AKANMORI Barthoromew D. Research Fellow, Noguchi Memorial Institute for Medical Research, University of
POLJAK Robe パスツール研究所, 兔疫構造部門, 教授
|Project Period (FY)
1989 – 1991
Completed(Fiscal Year 1991)
|Budget Amount *help
¥10,100,000 (Direct Cost : ¥10,100,000)
Fiscal Year 1991 : ¥3,300,000 (Direct Cost : ¥3,300,000)
Fiscal Year 1990 : ¥3,300,000 (Direct Cost : ¥3,300,000)
Fiscal Year 1989 : ¥3,500,000 (Direct Cost : ¥3,500,000)
|Keywords||Malaria vaccine / X-ray structural studies / Repeating amino acid sequence / Circumsporozoite surface protein as antigen / Monoclonal antibodies against circumsporozoite / surface protein / Amino acid Sequence of heavy chain in Fab / 融合蛋白質抗原 / モノクロ-ナル抗体 / 抗原・抗体複合体|
Malaria is a Darasitic disease transmitted by mosquitoes and a major cause of death in most tropical areas of the world, because of the appearance of insecticide-resistant mosquitoes and drug-resistant malaria parasites. These have prompted the search for other alternative methods to eradicate it. The objective of this research project was to develop a malaria vaccine which wbold. attack the invaded sporozoite parasite. In order to achieve this purpose, we finished the following subjects in the fiscal years (1989 - 1991).
1) Expression and purification of the fusion protein (FP) consisting of a part of human growth hormone and an amino acids-repeating segment, (PNAN)_8(PNVD)_2, of circumsporozoite surface protein (CSP) in E. coli, and the CD measurement or postulation of its secondary structure.
2) Purification of the monoclonal antibodies (Ab) against FP produced in BALb/c mice, and isoldrion of Fab segment (Ab-Fab) by partial digestion with papain enzyme.
3) ELISA tests for affinity of the antigen (FP) for antibody (Ab) were performed, and the dissociation constants were experimentally determined.
4) Crystallization trials of the antigen (FP), the antibody (Ab-Fab) and the complex of AbFab with FP. by using several precipitants under different temperature and pHs have been started, and the antibody crystal grew to 0.8 mm in two to three weeks, but they are still too small in dimension for X-ray data collection.
5) The amino acid sequence determination of antibody started in 1990 at the Institut Pasteur in Paris according to G. Winter's procedure. We completely determined the amino acid sequence of the heavy chain in Ab-Fab in 1991, and found two consensus cysteine residues in comparison with the other antibody sequences.
6) We succeeded in overproducing a new fusion protein (FP-l) consisting-of FP and a peptide which was found in CSP and acted. as a T-cell epitope and its-crystallization.