Generation and characterization of monoclonal antibodies to porcine zona pellucida-3 α glycoprotein

Abstract

All 10 monoclonal antibodies generated reacted with deglycosylated ZP3 alpha both in ELISA and Western blots, thereby suggesting that these do not recognize carbohydrate determinants. Moreover, 7 antibodies (MAs −402, −403, −405, −412, −420, −421 and −423) recognized reduced and carboxyamidomethylated ZP3 alpha in Western blot suggesting that these antibodies recognized linear epitopes. The epitopes recognized by MAs −410, −413 and −425 were sequential or conformational, stabilized by disulphide bonds. Three antibodies namely, MA −405, −420 and −421 inhibited in vitro, zona lysis by trypsin.