|
Phospholipase A,(PLA,) plays important roles in several biological phenomena such as mem-brane phospholipaid turnover, signal transduction through liberation of arachidonate form mem-brane phospholipids and the process of inflammation. In order to detect the unknown PLA,s in febrile patients urine, three kinds of monoclonal antibodies that react with PLA,s(Crotalus atrox and Naja naja atra PLA,) were established. Among thirty monoclonal antibodies obtained, results obtained with three typical monoclonal antibodies were characterized. KS-1 was highly specific to Naja naja atra PLA, and no cross-reaction with other PLA,s was observed using ELISA(enzyme linked immunosorbent assay). KS-1 significantly inhibited the hemolytic activity of Naja naja atra PLA,in a dose dependent manner. On the other hands, KS-2 was also highly specific to Crotalus atrox PLA but did not cross-react with other two PLA,s. Furthermore, KS-2 did not inhibit anticoagulant activity of Crotalus atrox PLA. Another distinct monoclonal an-tibody, designated NP-1, was further characterized. NP-1 respectively showed cross-reactivity with Crotalus atrox and human pleural PLA, on ELISA, and also significantly inhibited an- ticoagulant activity of Crotalus atrox PLA,. The results of epitopes analysis by the inhibition tests of anticoagulant activity and ELISA inhibition assay of PLA,s, NP-1 recognized only an-ticoagulant sites of three dimensional conformation of Crotalus atrox PLA
|