Factors affecting the quality of cytochrome C prepared by biological separation.

Cytochrome C is a biochemical drug widely used in clinical treatment. To improve the quality of cytochrome C, we studied the factors influencing the quality of cytochrome C in the preparation process. The purity of cytochrome C prepared by different processes was analyzed by SDS-PAGE, and the 2. 5x10⁴ protein in SDS-PAGE was analyzed and identified by LC-MS/MS. The results of SDS-PAGE electrophoresis show that the sample of cytochrome C prepared by traditional process using trichloroacetic acid (TCA) precipitator contained not only 1. 22 x 10⁴ cytochrome C, but also a protein molecule of about 2. 5 x 10⁴. Cytochrome C samples prepared by the new process without trichloroacetic acid precipitator only had 1.22 x 10⁴ cytochrome C, but no 2. 5 x 10⁴ protein was found. The LC-MS/MS analysis and identification result of the 2. 5 x 10⁴ protein in the SDS-PAGE map show that the coverage between this protein and cytochrome C (susscrofa) was 76%. Combining with its molecular weight, it can be proved that this protein is cytochrome C dimer. TCA precipitator during the preparation of cytochrome C easily leads to the formation of cytochrome C dimer. Therefore, TCA is one of the key factors affecting the quality of cytochrome C. [ABSTRACT FROM AUTHOR]