Center of Genetic Engineering and Biotechnology, C. Havana, Cuba.
Biotechnol Bioeng. 1993 Nov 20;42(10):1238-44. doi: 10.1002/bit.260421014.
Recombinant hepatitis B surface antigen (r-HBsAg) produced in yeast is adsorbed on a diatomaceous earth matrix for purification purposes. A pH dependence in the adsorption-elution behavior was found. The capacity of celite (Hyflo Super Cei) for adsorbing r-HBsAg increased with decreasing pH. Nonspecific proteins were also adsorbed, but a low pH dependence was found. Elution from the matrix was performed using a basic pH buffer, in which r-HBsAg is more specifically adsorbed/desorbed than contaminant proteins, permitting the purification of the r-HBsAg. A pH of 4.0 was used for adsorption and pH 8.2 was used for desorption. The described protocol allows a purification factor between three- and fivefold with respect to contaminant proteins and sixfold with respect to contaminant DNA. Finally, the adsorption step was successfully scaled-up for production purposes.
酵母表达的重组乙型肝炎表面抗原（r-HBsAg）通过吸附到硅藻土基质上进行纯化。研究发现，该吸附洗脱行为存在 pH 值依赖性。Celite（Hyflo Super Cei）吸附 r-HBsAg 的能力随 pH 值降低而增加。非特异性蛋白质也被吸附，但发现其吸附的 pH 值依赖性较低。用碱性 pH 值缓冲液从基质中洗脱，r-HBsAg 比污染物蛋白更特异的被吸附/解吸，从而可以纯化 r-HBsAg。吸附时 pH 值为 4.0，洗脱时 pH 值为 8.2。该方法相对于污染物蛋白可获得 3 至 5 倍的纯化倍数，相对于污染物 DNA 可获得 6 倍的纯化倍数。最后，成功地将吸附步骤放大用于生产。
