磁性免疫微球在人血清白蛋白纯化中的应用(英文)

Rapid purification of human albumin with immunomagnetic microsphere technology

目的为了快速地从人血清中提纯人血清白蛋白,利用磁性免疫微球作为提取手段,再用间接酶联免疫法测定人血清白蛋白的回收率。方法将经过羧基修饰的聚苯乙烯微球作为载体,用EDC(碳化亚胺)活化微球表面的羧基,再将兔抗人血清白蛋白抗体包被于微球上,这种微球-抗体复合物能特异性地捕获人血清白蛋白,磁分离复合物后,通过将兔抗人血清白蛋白抗体作为捕获抗体,将酶联羊抗人血清白蛋白抗体作为检测抗体,建立起间接酶联免疫法,用于检测人血清中和磁性免疫微球上吸附人血清白蛋白的浓度,得到微球从人血清中提纯人血清白蛋白的回收率。结果第1次提纯的回收率为(86±4)%,重复利用微球2次,回收率分别为(69.0±0.6)%和(40.8±0.8)%,而提纯的人血清白蛋白的纯度为90%。结论以上结果表明,免疫磁性微球提纯人血清白蛋白的实验是有效的,为工业上大规模提纯人血清白蛋白提供了一条新的思路。

Aim To develop a rapid and inexpensive method for purification of human albumin, a method of immunomagnetic microspheres （IMMS） based on enzyme-linked immunosorbent assay （ELISA）for the purification of human albumin from human serum. Methods Polystyrene magnetic microspheres with carboxyl groups as carriers were prepared, and then the carboxyl groups on the surface of the microspheres were activated by ethylcarbodiimide （EDC）. Finally rabbit anti-human serum albumin （HSA） antibodies were covalently bound to it and the complex can specifically capture HSA. After the procedure of capturing HSA, through taking rabbit anti-human albumin protein antibodies as a capture antibody, and goat anti-human albumin protein antibodies as a detection antibody, an ELISA on IMMS was developed, which can determine the recovery yield of HSA from the human serum. Results The result of the experiment was that the recovery of human albumin with IMMS was （86 ± 4） % , and IMMS were reused for two other purifying cycles, the results of which were （69.0 ± 0.6） % and （40.8 ± 0.8） % , and the purity of the product was about 90%. Conclusion The results above prove that the immunomagnetic purifiying strategy was shown to be efficient and offers an new thought for a large scale production of highpurity HSA.