大肠埃希菌周质抗体细胞释放工艺方法研究

本文应用细胞裂解液提取法、超声波细胞破碎法、细胞冻融法、bugbuster提取法和高压细胞破碎法，对比研究了从大肠埃希菌周质中提取可溶性抗体的相关参数，并通过SDS-PAGE电泳，BCA蛋白浓度测定和ELISA法检测了上述四种方法的有效性。结果表明，对于试验室小试规模的抗CEA抗体的细胞释放，超声法破碎，提取的目的抗体量最多，优于细胞裂解液提取法，冻融法和bugbuster法，最佳条件为：细胞缓冲液与湿菌体的比例为

Research for extraction of periplasmic proteins in Escherichia coli

This article reviews several cell breakage methods,such as extraction buffer method,ultrasonic disruption,repetitive freezing and thawing,bugbuster and high-pressure cracker.Compare the periplasmic protein we get from each mehod,in aspects of weight and activity,by using SDS-PAGE electrophoresis,BCA Protein Assay Kit,and ELISA.The conclusions are as follows:in a small scale laboratory,ultrasonic disruption is the most efficacious method,and the optimal condition is putout power 250W,radiation/intermission time 4s/5s,total working times is 140,and the amount of cell buffer 5mL/g with ice bath.Besides,for the large-scale periplasmic protein preparation,the optimal method is high-pressure cracker in 850Pa,crushing twice.