[Construction and identification for cell strain of anti-human seminal plasma phospholipase A2 monoclonal antibody].
OBJECTIVE
To establish and evaluate the anti-human seminal plasma phospholipase A2 (PLA2) monoclonal antibody (McAb).


METHODS
After having been separated and purified from human seminal plasma by PEG precipitation, Sephacryl S-300 column chromatography, DEAE-Sephadex A-25 column chromatography and HA column chromatography, PLA2 was regarded as an antigen to immune BALB/C mouse to produce anti-human seminal plasma PLA2 McAb. The PLA2 McAb sensitivity and specificity were performed by ELISA technique and Western-blot analysis, respectively.


RESULTS
The molecular weight of PLA2 depurated with 245 fold purification from human seminal plasma was about 34,900, while the sensitivity and typing of its McAb were 1:5(6)-1:5(8) and IgM (kappa) with a satisfied Western-Blot results.


CONCLUSIONS
The PLA2, which had not been reported in international and domestic papers, may be a new type of PLA2. The establish of its McAb will provide significant tools for the research of the relationship between PLA2 in human seminal plasma and male fertility.
Wang
Shu-Kui
SK
Laboratory of Reproduction & Genetics, Nanjing General Hospital of Nanjing Command, PLA, Nanjing, Jiangsu 210002, China.
Huang
Yu-Feng
YF
Xia
Xing-Yi
XY
Li
Bao-Tong
BT
Wang
Zi-Zheng
ZZ
Liu
Meng-Liu
ML
D000911:Antibodies, Monoclonal / Q000276:immunology*; D003637:DEAE-Dextran / Q000031:analogs & derivatives* / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D008970:Molecular Weight; D010741:Phospholipases A / Q000276:immunology / Q000302:isolation & purification* / Q000378:metabolism; D054467:Phospholipases A2; D012661:Semen / Q000201:enzymology*
D000911:Antibodies, Monoclonal; C030237:DEAE-sephadex A-25; D003637:DEAE-Dextran; D010741:Phospholipases A; D054467:Phospholipases A2