Determination of specific antibodies in the serum of rabbits sensitized by 1,5-dicaffeoylquinic acid in Shuanghuanglian Injection
Objective To determine the specific antibodies in the serum of rabbits sensitized by 1,5-di caffeoylquinic acid in Shuanghuanglian Injection by ELISA.Methods The suspected small molecule hapten substance 1,5-dicaffeoylquinic acid in Shuanghuanglian Injection was incubated with normal rabbit serum in vitro to prepare a complete antigen,which was then used to immunize rabbits to obtain specific antibodies.The antibody titer of antiserum was detected by ELISA method.The serum specificity was determined by indirect competitive ELISA and the inhibition curve was plotted.The content of IgE antibodies in the antiserum of rabbits in each group was detected using a rabbit Immunoglobulin E(IgE)ELISA kit.Results The A values of 1,5-dicaf-feoylquinic acid of 0.25 mg/ml,0.50 mg/ml,1.00 mg/ml,2.00 mg/ml,and 4.00 mg/ml were(0.364±0.020),(0.334±0.010),(0.322±0.020),(0.332±0.010),and(0.298±0.020),respectively,which were higher than the A value of the negative control group(0.095±0.010).The regression equation obtained was I=0.195 2 lgC+0.273 4,with R2=0.995 8,and the half maximal inhibitory concentration(IC50)was 33.494 mg/ml.Exogenous IgE-type antibodies were obviously produced in rabbits.Conclusion The suspected hapten 1,5-dicaffeoylquinic acid in Shuanghuan-glian Injection is allergenic under the experimental conditions.
1,5-dicaffeoyl quinic acidShuang Huang Lian Zhu She JiEnzyme-linked immunosorbent assay
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