PREPARATION OF A STABLE NON-INFECTIVE COMPLEMENT-FIXING ANTIGEN FOR HERPES SIMPLEX

Abstract

The soluble antigen of herpes simplex is prepared by grinding the specifically infected chorioallantoic membranes with one volume of saline, storage for 4 days at 4 °C to precipitate tissue material, followed by centrifugation for 1 hour at 10,000 r.p.m. (9000 g) to improve the specificity. Nine different methods of formaldehyde treatment were tried to determine the best procedure for the destruction of the infectivity while preserving the maximal antigenicity. Methods in which treatment was conducted at pH 6 destroyed considerable antigenicity whereas the methods using a pH of 8.5 were most successful. The method selected to inactivate the virus present in the herpes antigen involves treatment with 0.01% formaldehyde at 37 °C for 2 days at pH 8.5, followed by the addition of 0.25 cc of 30% dibasic ammonium phosphate solution per 100 cc of treated antigen. After treatment and neutralization the antigen can be lyophilized for stable storage.