The research of a SEC-UV-RI method to determine the modifiction degree of PEG-rhGH
10.16438/j.0513-4870.2024-0576
- VernacularTitle:双检测器串联测定聚乙二醇化人生长激素平均修饰率方法研究
- Author:
Yi LI
;
Xiao-ming ZHANG
;
Xin-yue HU
;
Yue SUN
;
Lü-yin WANG
;
Du-yu CHEN
;
Ping LÜ
;
Cheng-gang LIANG
;
Jing LI
- Publication Type:Research Article
- Keywords:
size exclusion chromatograph;
ultraviolet and refractive index detection in series;
PEGylation;
growth hormone;
average degree of modification
- From:
Acta Pharmaceutica Sinica
2024;59(12):3367-3373
- CountryChina
- Language:Chinese
-
Abstract:
According to the requirements of the regulatory authorities, degree of modification (DP) should be included in the characterisation of the PEGylated protein drug substance, and is one of the critical quality attributes for quality control. In this study, based on the fundamental assumption that the refractive index (RI) signal and the ultraviolet (UV) signal of PEGylated protein are equal to the sum of the corresponding signal produced by the polyethylene glycol (PEG) and protein parts of the conjugates in their uncoupled state, we developed a method to determine the DP of PEGylated recombinant human growth hormone (inpegsomatropin). In this method, 20 μL of 1 mg·mL-1 human growth hormone (hGH) standard, 2 mg·mL-1 PEG reference substance and 1 mg·mL-1 drug substance solution were each injected to size exclusion chromatographic (SEC) column for separation, detected with ultraviolet and refractive index (UV-RI) detectors in series. Finally, the DP was calculated as the formula derived from the fundamental assumption. The developed SEC-UV-RI method showed good specificity, repeatability (RSD = 0.63%, n = 9) and accuracy, with a recovery of 100.0%, compared with the result obtained from the simultaneously established classical acid hydrolysis method, demonstrating pretty handleability and accessibility, and is appropriate for quality control test of DP for this drug substance.
