1991 Fiscal Year Final Research Report Summary
Isolation and characterization of an novel oncodevelopmentalprotein from human malignant ascitic fluid.
| Project/Area Number |
01570939
|
|---|
| Research Category |
Grant-in-Aid for General Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Research Field |
Obstetrics and gynecology
|
|---|
| Research Institution | Kitasato University School of Medicine |
|---|
Principal Investigator |
NAKAI Mitsuo Kitasato University School of Medicin, Dept. of OB/GYN., Professor, 医学部, 教授 (50050399)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
IINO Joji 北里大学, 医学部, 助手 (30176052)
ISHIKAWA Masakazu 北里大学, 医学部, 助手 (40184494)
YAMAURA Noboru 北里大学, 看護学部, 助教授 (60050621)
|
|---|
| Project Period (FY) |
1989 – 1991
|
| Keywords | Tumor-associated protein / Oncedevelopmental protein / Pregnancy-associated protein / Tumor marker / Isolation / Characterization / Serum concentration / Gynecological malignant diseases |
|---|
| Research Abstract |
A tumor-associated antigen that shares antigenicity with a pregnancy-associated protein has been detected in ascitic fluid of patients with advanced ovarian can cer. The protein was prepared from malignant ascitic fluid by the combination of poly (ethylene glycol) 4000 fractionation, DEAE-Sepharose column chromatography and prepardtive polyacrylamide gel electrophresis (PAGE). Antiserum against an malignant ascitic antigen (MASA) was raised in rabbits. The rabbit antiserum was applied to a column of normal human plasma (NHP)-coupled Sepharose, and specific antibodies against MASA (anti-MAsA) was highly purified. Subsequently, by the use of anti-MAsA-coupled Sepharose, highly purified MASA was obtained. The preparation thus obtained was analyzed on sodium dodecyl sulfate (SDS)-PAGE, Using this technique, a single antigenically active component was identified" The moleculermass of this MASA species was calculated to be 27 kDa by SDS-PAGE.
Highly sensitive assay method employed ELISA was established by the use of the monospecific anti-MAsA. With this methody- the smallest detectable amount detectable in a sample was 0.00125 U/ml. The sensitivity 6f the ELISA is 1, 000-fold higher than that of a previous electroimmunoassay employed a modified Laurell's method. With this method, it will be possible to establish the normal level range, which is a prerequisite for precise evaluation of the increase in abnormal conditions. In conclusion, the present study shows that estimation of the MASA will be an usefulladfunct for the management of patients with gynecological malignant diseases.
|
