Method development and validation for testing the concentration of anti-TNF-α monoclonal antibody in serum based on ELISA
10.13699/j.cnki.1001-6821.2024.11.021
- VernacularTitle:用ELISA法检测血清中抗TNF-α单克隆抗体药物浓度的方法开发与验证
- Author:
Zhen-Xiang HU
1
;
Li-Xiu HE
;
Bo WANG
;
Xi CHEN
;
Gui-Li LIU
;
Yu-Min QIN
Author Information
1. 珠海市丽珠单抗生物技术有限公司,广东珠海 519090
- Keywords:
tumor necrosis factor-α;
enzyme linked immunosorbent assay;
monoclonal antibody;
serum;
drug concentration
- From:
The Chinese Journal of Clinical Pharmacology
2024;40(11):1642-1645
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish an indirect enzyme-linked immunosorbent assay(ELISA)method for testing the concentration of a monoclonal antibody target tumor necrosis factor-α(TNF-α)in animal serum.Methods The critical parameters of the method including coating concentration of human TNF-α,source,concentration and stability of HRP-labeled goat anti-human immunoglobulin G(IgG)were investigated.The specificity,accuracy,precision,linearity and Limited of Determination of the method were investigated.Results The critical parameters of the method were confirmed as below:TNF-α was coated at 400 ng·mL-1;HRP labeled goat anti-human IgG antibody was diluted at 1:3.0 ×105;the diluted horseradish peroxidase labeled goat anti-human IgG antibody is well stored at 4 ℃ for 3 days.Meanwhile the method was confirmed to have good specificity,the recovery rate ranged from 84.00％to 106.82％,the coefficient of variation of different antibody concentration levels were no more than 10％;the method had a good linearity and the standard curve was y=(-8.37×103-2.37 × 106)/[1+(x/29.80)106]+2.37 × 106(R2=0.999);the limit of quantification was 1 ng·mL-1,all of which met the requirements.Conclusion A accurate and robust ELISA method was developed to test the concentration of anti-TNF-α monoclonal antibody in serum.
