RECOVERY PROCESS OF PROTEIN INCLUSION BODIES RECOMBINANTLY PRODUCED BY E. COLI.

摘要

The protein inclusion bodies expressed in E. coli is purified by centrifuging E. coli culture media at low temp., collecting its precipitate, suspending the ppt. in an acidic buffer which is containing an surfactant in order to suppress unstable factors, homogenizing, centrifuging at low temp., and re-collecting the ppt.(homogenizing step); suspending and strring the ppt. in a solution which is composed of 0.5-0.1% nonionic surfactant and 10-50mM EDTA, diluting with a solution contg. 0.05-0.2% surfactant and 100-500mM NaCl, centrifuging, and collecting the ppt. which is containing the protein inclusion bodies removed almost all impurities e.g. cell membranes etc.(NaCl washing step); finally resuspending the inclusion bodies in an adequate amount of distilled water, stirring violently, centrifuging, and collecting the highly pure inclusion bodies(washing step with distilled water).