重组蛋白A免疫吸附剂的制备及吸附性能研究

Preparation of a recombinat protein A-based adsorbent and its adsorption properties

以琼脂糖凝胶为载体,重组蛋白A为配基制备免疫吸附剂。过血浆动态吸附之后洗脱得到含有免疫球蛋白的洗脱液;通过分析洗脱液中免疫球蛋白的纯度可以考察吸附剂对免疫球蛋白的吸附选择性;最后利用等温静态吸附检测了吸附剂对人免疫球蛋白(hIgG)溶液的吸附行为。结果表明,洗脱液中免疫球蛋白的纯度大于92%,重组蛋白A吸附剂对免疫球蛋白吸附选择性较好;等温静态吸附结果符合Langmuir等温吸附模型;进一步分析Langmuir吸附模型,表明该吸附剂在临床治疗中有吸附量大的明显优势,重组蛋白A免疫吸附剂具有很好的临床应用前景。

An adsorbent has been prepared by attaching recombinant protein A as the ligand to Sepharose 6FF as the support.The adsorption selectivity of the adsorbent was assessed by analyzing the abundance of eluted human immunoglobulin after dynamic adsorption of plasma.The adsorption behavior of the adsorbent was tested by isothermal adsorption of human immunoglobulin solution.It was found that the purity of human immunoglobulin in the eluant was more than 92%,suggesting that the recombinant protein A adsorbent has a strong preference for immunoglobulin.The results of isothermal adsorption are in agreement with the Langmuir isotherm equation with a dissociation constant of 0.36 mg/mL and a maximum capacity of 35.7 mg/mL.Our study demonstrates that the recombinant protein A-based adsorbent has good affinity for immunoglobulin and has potential clinical applications.