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Translation products of pre-S(1), pre-S(2) regions and the S gene of hepatitis B virus: susceptibility of their antigenic activities to treatment with heat, urea formalin or pepsin
Authors:H. Ohnuma   A. Machida   Emiko Takai   F. Tsuda   H. Miyamoto   T. Tanaka   K. Oda   S. Usuda   T. Nakamura   Y. Miyakawa     M. Mayumi
Abstract:Hepatitis B subviral particles, purified from plasma of asymptomatic carriers seropositive for hepatitis B e antigen, were treated with various conditions reported for the processing of vaccines. Thereafter, antigenic activities displayed by the translation products of pre-S(1), pre-(2) regions and the S gene were determined with monoclonal antibodies, and the reactivity for polyalbumin receptor was tested. Heating at 100 degrees C for 1.5 min and then at 65 degrees C for 10 h preserved more than 1/2 of antigenic activities representing products of pre-S(1), pre-S(2) regions and the S gene. After incubation in the presence of 8 M urea at 37 degrees C for 4 h, more than 2/3 of antigenic activities still remained. The antigenic activity of the S gene product was decreased to 2/3 and that of pre-S(2) region product to 1/3, after treatment with formalin at the final concentration of 1:4000 at 37 degrees C for 72 h, whereas the activity of pre-S(1) region product was affected drastically. Although 1/5 of the antigenic activity of the S gene product survived the digestion with pepsin for 18 h, antigenic activities of pre-S(1) and pre-S(2) region products were destroyed almost completely. Polyalbumin receptor, borne by the pre-S(2) region product, was lost by pepsin digestion also. Based on the results obtained, heating may be most appropriate for sterilizing plasma-derived hepatitis B particles for use as a vaccine, because it is reliably virucidal and would not affect the protective efficacy to an extent as the other virucidal methods would.
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