Immunological properties of recombinant hepatitis B surface antigen expressed in mammalian cell (C127).
We examined the immunological properties of the recombinant hepatitis B surface antigen (r-HBsAg) which was expressed in mammalian cell (C127). The cross-immunity of r-HBsAg and plasma-derived hepatitis B surface antigen (p-HBsAg) were tested using Western blotting and ELISA with guinea pig polyclonal antibody and naturally infected human-derived antibody and the both antigens show the same results in their response pattern and intensity, which indicate they have a good cross-immunity. From the measurement of ED50 after formalin- or heat-inactivation, both r-HBsAg and p-HBsAg showed ED50 of 0.2-0.3 in formalin-inactivation, while r-HBsAg was 0.05-0.09 and p-HBsAg was 0.03-0.07 in heat-inactivation, which means heat-inactivation method is 3-4 times superior in immunogenicity. In the immunopersistency test performed in guinea pig for the period of 3 months with two different adjuvants, antibody titer was 34.2 with muramyl dipeptide adjuvant, which was 1.8 times greater than the antibody titer of 18.9 with AlPO4 adjuvant. The mutagenicity of r-HBsAg, examined by Ames test and micronucleus test, were all negative. In conclusion, r-HBsAg has the same cross-immunity with p-HBsAg, and heat-inactivation method and muramyl dipeptide adjuvant allow development of r-HBsAg vaccine with excellent immunogenicity.
Lee
Y S
YS
College of Pharmacy, Seoul National University, Kwanak-Gu, Korea.
Kim
B K
BK
Choi
E C
EC
D000276:Adjuvants, Immunologic; D000818:Animals; D000918:Antibody Specificity; D015153:Blotting, Western; D002478:Cells, Cultured; D004797:Enzyme-Linked Immunosorbent Assay; D005557:Formaldehyde / Q000737:chemistry; D006168:Guinea Pigs; D006514:Hepatitis B Surface Antigens / Q000096:biosynthesis / Q000737:chemistry / Q000276:immunology*; D006358:Hot Temperature; D007111:Immunity, Cellular / Q000276:immunology; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000276:immunology; D013154:Spleen / Q000166:cytology
D000276:Adjuvants, Immunologic; D006514:Hepatitis B Surface Antigens; D011994:Recombinant Proteins; D005557:Formaldehyde