蛋白质快速定量法及其在rhZP3纯化中的应用  

A rapid method for protein quantification and its application in rhZP3 purification

作　　者：谢琪璇[1] 郑育声[2] 彭雅林[1] 肖銮娟[1] 余巍[1] 潘善培[1] 

机构地区：[1]暨南大学生殖免疫研究中心,广州510632 [2]海南大学生物工程系,海口570002

出　　处：《第三军医大学学报》2005年第12期1208-1210,共3页Journal of Third Military Medical University

基　　金：国家自然科学基金资助项目 ( 30 370 54 1) ;广东省自然科学基金资助项目 ( 31883) ;广东省科技计划资助项目 ( 2 0 0 3830 50 1)~~

摘　　要：目的　建立层析过程中洗脱峰蛋白质含量的快速定量法。方法　利用蛋白质的紫外吸收原理和层析软件的积分功能,即A2 80 和峰面积计算层析过程中洗脱峰蛋白质含量。结果　洗脱峰大小与蛋白质含量呈线性关系,洗脱峰蛋白质含量可用方程X =Y A进行估算,X为待测蛋白含量(mg) ,Y为洗脱峰面积(mAU×ml) ,A为1g L待测蛋白在层析系统的紫外吸收值(mAU)。将其应用于重组人卵透明带ZP3蛋白(rhZP3 )的纯化,用该方程估算的纯化蛋白含量与用Low ry法测定结果相似。结论　该法简便、快速、重复性好,具有一定实用价值。Objective To find a rapid and simple method for quantifying the amount of eluted proteins in chromatography. Methods The method was developed by utilizing the principle of the absorbance of protein at 280 nm and the integral function of the software UNICORN for AKTA FPLC, namely, the protein quantity was evaluated based on its A_ 280 and peak area. Results The relation between peak area and quantity of the protein was linear correlation. The protein quantity could be expressed as X=Y/A. The Y represented the peak area (mAU×ml) of the interested fraction, and A represented the measurable value (mAU) of 1 g/L interested protein through the FPLC 280 nm cell and X was the quantity of the interested protein (mg). The method was used in the purification of recombinant human zona pellucida-3 protein (rhZP3)and the quantity of the purified rhZP3 evaluated from the formula was confirmed by Lowry method. Conclusion The method is easy, rapid, repeatable and practicable to quantify the protein in chromatography.

关 键 词：层析 蛋白 定量 重组人卵透明带ZP3蛋白 纯化 

分 类 号：Q503[生物学—生物化学] Q513