ABSTRACT
OBJECTIVE To investigate the immunogenicities of Poria cocos polysaccharides, PCP-Ⅰand PCP-Ⅱ, as a vaccine adjuvant. METHODS ①Keyhole limpet hemocyanin (KLH) was linked to PCP-Ⅰor PCP-Ⅱrespectively to prepare immuno-antigen KLH-PCP-Ⅰor KLH-PCP-Ⅱ. Bovine serum albumin (BSA) was also linked to PCP-Ⅰor PCP-Ⅱrespectively to prepare screening-antigen. Rabbits were immunized with KLH-PCP-Ⅰor KLH-PCP-Ⅱplus Freund adjuvant by intradermal injection twice, and serum specific antibody titers were determined by ELISA. ②BALB/c mice were immunized with PCP-Ⅰ or PCP-Ⅱ alone intramuscularly twice, and serum polysaccharide antibody titers were determined by ELISA.③BALB/c mice were co-immunized intramuscularly or subcutaneously with PCP-Ⅰor PCP-Ⅱplus hepatitis B surface antigen (HBsAg) or porcine reproductive and respiratory syndrome virus inactivated vaccine (PRRSV) twice, and serum polysaccharide-antibody titers were determined by ELISA. RESULTS ①Serum anti-KLH and anti-polysaccharides (PCP-Ⅰor PCP-Ⅱ) antibodies were pro?duced after rabbits were immunized with KLH-PCP-Ⅰor KLH-PCP-Ⅱplus Freund adjuvant twice.②Serum anti-PCP-Ⅰor anti-PCP-Ⅱantibodies were not found after mice were immunized with PCP-Ⅰand PCP-Ⅱalone twice.③After mice were immunized with HBsAg or PRRSV plus PCP-Ⅰor PCP-Ⅱtwice, serum anti-PCP-Ⅰor anti-PCP-Ⅱantibodies were not found. CONCLUSION PCP-Ⅰand PCP-Ⅱshow weak immunogenicity, which may be quite safe as a vaccine adjuvant.
ABSTRACT
Objective To develop a sandwich enzyme-linked immunosorbent assay ( ELISA) for the determination of a humanized antibody MIL50.Methods RiVax, a mutant of RTA (a chain of ricin) expressed in E.coli, was used as the coating protein.Horseradish peroxidase (HRP) labeled IgG of goats against humans was used to determine MIL 50 captured by RiVax coated on the plate.Results Compared with ricin, RiVax could bind well with MIL50,indicating it could be used as a coating protein to determine MIL50 instead of holotoxin.Using this method, the detective limit of MIL50 in PBST (PBS containing 0.1%Tween 20) could be as low as 0.030 mg/L.The absorbance value of ELISA for MIL50 in the ser-um and homogenate of the liver , spleen, lung, kidney, muscle of rats was lower than that in PBST .Conclusion The sandwich ELISA is a sensitive method for the analysis of distribution of MIL 50 in rat tissues.