Purification, Characterization of Seminal Fibronectin Fragment in Human Seminal Plasma and Its Apllication for Legal and Clinical Medicine

1997 Fiscal Year Final Research Report Summary

• Project/Area Number: 08670492
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Legal medicine
• Research Institution: Nagasaki University
• Principal Investigator: TSUDA Ryouichi Nagasaki University, School of Medicine, Lecture, 医学部, 講師 (20098875)
• Co-Investigator(Kenkyū-buntansha): NAKASONO Ichiro Nagasaki University, School of Medicine, Professor, 医学部, 教授 (30108287) ; KUBO Shin-ichi Tokusima University, School of Medicine, Professor, 医学部, 教授 (10205122)
• Project Period (FY): 1996 – 1997
• Keywords: Fibronectin / Seminal plasam / Idenntification / seminal specific antigen / Immunohistochemistry / Forensic immunology

Research Abstract

A 36kDa protein was isolated and purified by affinity chromatography utilizing blue-sepharose, gel filtration on sephadex G-100 and Fast Protein Liquid Chromatography System from human seminal plasma. The protein have a single band in 12% SDS-polyacrylamide gel electrophoresis. The purified protein sequenced by automated Edman degradation. The N-terminal sequence of the protein was found to corresponded exactly with the sequence of residues 1230 to 1237 at cell attachment domain of human fibronectin. The fact suggest the 36kDa protein was fibronectin fragment. The anti-seminal fibronectin fragment serum was prepared by immunizing rabbit the purified protein emulsified with complete Freund's adjuvant. In human normal seminal plasma (n-54), concentration range of the seminal fibronectin fragment was 316-2,190 ug/ml (mean-996 ug/ml). Cross-reaction was found the purified plasma fibronectin by ELISA using anti-seminal fibronectin fragment serum. By ELISA and immunohistochemical method, the localization of seminal fibronectin flagment in male genital organs was detected in seminal vesicle.