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目的制备丹酚酸B(SAB)的人工抗原(SAB-BSA)和包被原(SAB-PLL)并进行鉴定。方法采用碳二亚胺(EDC)法将SAB分别与牛血清白蛋白(BSA)和多聚赖氨酸(PLL)载体偶联合成SAB-BSA和SAB-PLL;采用紫外分光光度(UV)法和薄层色谱(TLC)法对偶联效果进行鉴定,并通过SAB与BSA的吸光值计算出二者的偶联比;利用获得的SABBSA免疫小鼠,采用酶联免疫吸附法(ELISA)检测小鼠抗血清内多克隆抗体效价并以间接竞争酶联免疫吸附法(ic-ELISA)检测小鼠抗血清内多克隆抗体的灵敏度。结果所合成的SAB-BSA为BSA和SAB的结合物,且SAB-BSA中无游离SAB,SAB与BSA偶联成功,SAB-BSA偶联结合比为18:1;SAB-BSA刺激小鼠产生的针对SAB的多克隆抗体,效价为1:2×10~3,该多克隆抗体IC_(50)为17.7μg/mL。结论 SAB-BSA和SAB-PLL合成成功,可用于SAB单克隆抗体的制备以及SAB快速检测试剂盒的研制。
OBJECTIVE To prepare and identify the artificial antigen (SAB-BSA) and envelop (SAB-PLL) of salvianolic acid B (SAB). Methods SAB-BSA and SAB-PLL were synthesized by coupling carbazole (EDC) with bovine serum albumin (BSA) and polylysine (PLL) carrier, respectively. UV spectrophotometry And thin layer chromatography (TLC) were used to identify the coupling effect. The coupling ratio was calculated by the absorbance values of SAB and BSA. The mice were immunized with SABBSA, and the enzyme-linked immunosorbent assay (ELISA) The titer of polyclonal antibody in murine antiserum was determined by indirect competitive enzyme-linked immunosorbent assay (ic-ELISA). Results The synthesized SAB-BSA was a combination of BSA and SAB, and there was no free SAB in SAB-BSA. SAB-BSA conjugation was successful with SAB-BSA coupling ratio of 18: 1. SAB-BSA stimulated mice to produce Of polyclonal antibody against SAB with a titer of 1: 2 × 10 -3 and a polyclonal antibody IC 50 of 17.7 μg / mL. Conclusion SAB-BSA and SAB-PLL were successfully synthesized and could be used in the preparation of SAB monoclonal antibody and SAB rapid detection kit.