A study of non-reduced SDS-PAGE purity method of conbercept
10.16438/j.0513-4870.2018-0555
- VernacularTitle:康柏西普非还原SDS-PAGE纯度方法研究
- Author:
Chuan-fei YU
1
;
Feng ZHANG
1
;
Ai-bing LIU
2
;
Ya-fei ZHANG
2
;
Zu-xiu LUO
2
;
Su CHEN
2
;
Xiao KE
2
;
Lan WANG
1
Author Information
1. National Institutes for Food and Drug Control, Beijing 100050, China
2. Chengdu Kanghong Pharmaceutical Group, Chengdu 610000, China
- Publication Type:ORIGINAL ARTICLES
- Keywords:
non-reduced SDS-PAGE;
linearity;
loading amount
- From:
Acta Pharmaceutica Sinica
2018;53(12):2099-2103
- CountryChina
- Language:Chinese
-
Abstract:
A non-reduced SDS-PAGE purity method for quantitation of conbercept fragments was established based on gel screening, comparison of gel imaging system, linearity range of main band, screening of destaining conditions. The results indicated that the bands could be separated effectively with good clearness and flatness on 4%-15% gradient concentration gel, the peaks of all bands could be separated from baseline using high-distinguishability gel imaging system, the signal intensity of a main band had shown a good linearity with ≤ 3 μg of loading amount, and that the destaining was set as a total of ≤ 3 h with exchanging 100 mL destaining buffer every 60 min. The established non-reduced SDS-PAGE method could demonstrate the purity of conbercept more objectively. After validation, the established non-reduced SDS-PAGE method was submitted to FDA in the form of supplementary materials, which laid a quality basis for the direct entry of conbercept to the clinical Ⅲ study in the United States.
