Preparation of herpes simplex antigen for the diagnosis of carcinoma

A process for the industrial production of a non-virion antigen of the Herpes simplex virus for the diagnosis of the epithelial tissue tumors or carcinoma, in which the cells to be infected by the antigen are prepared starting from the guinea pig kidneys with the addition to said material of sterile trypsin and a culture medium consisting of lactoalbumin and calf serum, comprising the steps of:

• (i) growing the cells with a dynamic system inside Spinner flasks provided with magnetic revolving stirrers rotating at speeds higher than about 50 r.p.m. and operating at a temperature of 37.degree. C. and with successive additions of culture medium, inoculating the so obtained cells with Herpes simplex virus to with added culture medium, said infection phase being performed according to a volume ratio of the virus to the cells equal or higher than 1, but not higher than 100, the contact between the cells and the virus being maintained for about 3 hours;
• and collecting the antigen which is formed therein;
• (ii) purifying the collected antigen, by removing therefrom all the proteins and impurities which can negatively affect the tests for the diagnosis;
• comprising, a first centrifugation with recovery of the liquid phase from the first centrifugation step;
• precipitating the antigen protein and other proteins, a second centrifugation;
• diluting the residual solids from the second contrifugation using a buffer;
• purifying said diluted solids by molecular separation using organic gellified copolymers extraction column, collecting the proteins from the antigen purification on a column wherein the proteins are separated on the basis of their different electric charges;
• eluting said collected proteins and removing the non-glycosilated proteins by affinity chromatography on a column;
• and recovering purified antigens.