Verification and application of ELISA for determination of residual factor Ⅺ content in high concentration human immunoglobulin for intravenous injection
10.13200/j.cnki.cjb.004440
- VernacularTitle:高浓度静注人免疫球蛋白中人凝血因子Ⅺ残留量ELISA检测方法的验证及应用
- Author:
LI Zexiu
- Publication Type:Journal Article
- Keywords:
High concentration human immunoglobulin for intravenous injection(IVIG);
ELISA;
Human coagulation factor Ⅺ(FⅪ);
Residues
- From:
Chinese Journal of Biologicals
2025;38(03):352-358
- CountryChina
- Language:Chinese
-
Abstract:
Objective To verify and preliminarily apply the ELISA method for determination of human coagulation factor Ⅺ(F Ⅺ) residues in high concentration(10%) human immunoglobulin for intravenous injection(IVIG),so as to monitor the content of FⅪ in process samples.Methods The dilution reliability,accuracy,precision,limit of quantitation precision,linearity and range,and durability of the ELISA method for determination of FⅪ residues in 10% IVIG were verified.The FⅪ residues in three batches of 10% IVIG in the preparation process samples and the final products were determined by the verified method,and the results of the final products were compared with those of the commercially available IVIG(pH 4).Results Matrix effect existed in the determination of FXI residues in 10% IVIG by using this method,which could be solved by diluting 5-80 times.The average spike recovery rate of verification for accuracy was 100.1%.The relative standard deviations(RSDs) of repeatability verification by two experimenters were 8.54% and 5.70%,respectively,and the RSD of intermediate precision verification was 7.13%.The limit of quantitation concentration was 0.05 ng/mL,and the RSDs of limit of quantitation repeatability verification by two experimenters were 6.39% and 3.77%,respectively,and the RSD of intermediate precision verification was 12.00%.There was a good linear relationship between the concentration of F Ⅺ and A_(450) in the range of 0.01-5.00 ng/mL(r=0.999 9).Durability verification showed that different incubation time and 15 d storage under the specified conditions after opening the kit had no effect on F Ⅺ detection.The octanoic acid precipitation process removed F Ⅺ from 10% IVIG stably and effectively,and the residual amount of F Ⅺ in the three batches of 10% IVIG was lower than that in the commercially available IVIG(pH 4).Conclusion The method has good accuracy,precision and durability,and can satisfy the determination of FⅪ residues in 10% IVIG in laboratory.
