Determination of Specific Antibodies in Rabbit Serum Following Sensitiza-tion with Baicalein in ShuangHuangLian Injection
OBJECTIVE To detect the titer of specific antibodies in baicalin-sensitized rabbit serum of Shuang-HuangLian injection by enzyme-linked immunosorbent assay(ELISA).METHODS Normal rabbit serum was incu-bated with baicalin,a suspected small molecule hapten substance in ShuangHuangLian injection,in vitro to prepare the required complete antigens,and then the rabbits were immunized with these complete antigens in order to obtain specific antibodies.The antibody titer of the antiserum was detected by ELISA.By indirect competitive ELISA,serum specificity was further determined and the relevant concentration inhibition curves were plotted.In addition,rabbit im-munoglobulin E(IgE)ELISA kit was used to detect the corresponding IgE antibody content in the antiserum of rab-bits in each group.RESULTS The antibody titer(OD)value of baicalein was twice that of the negative control group,indicating its potential allergenicity.By collating the data and plotting the inhibition rate curve,the regression equations I=0.138lgC+0.402 and R2=0.959 were obtained,showing a good fit.The half inhibitory concentration was IC50=5.178 mg·mL-1.In addition,the results of this experiment clearly showed that exogenous IgE antibodies had been produced in rabbits,which further confirmed the sensitization of baicalin.CONCLUSION The results of the whole process of rabbit sensitization simulated in this laboratory showed that baicalein had obvious allergenici-ty.This finding has important guiding significance for the safe use of ShuangHuangLian injection,and also provides a valuable reference for the development and production of related drugs.
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