Shalini Swati, Sharma Anu, Mishra Nripendra Nath, Sharma Ratnesh Kumar, Chander Harish, Anvikar Anupkumar R, Chand Subhash
Therapeutic Antibody Laboratory, National Institute of Biologicals, (Ministry of Health and Family Welfare) Plot No. A-32, Sector-62, Institutional Area, Noida, Uttar Pradesh, 201309, India.
Heliyon. 2023 Feb 10;9(2):e13570. doi: 10.1016/j.heliyon.2023.e13570. eCollection 2023 Feb.
Vedolizumab is a humanized monoclonal antibody used for inflammatory bowel disease treatment. Vedolizumab binds to the α4β7 integrin complex and inhibits its binding to mucosal addressin cell adhesion molecule-1 (MAdCAM-1). To evaluate the binding efficacy and quality control check of Vedolizumab, flow cytometry is performed by using HuT78 cells. As we know, flow cytometer is costly and require high equipment maintenance with a designated technical manpower to handle it. In this regard, the aim of study was to develop and validate an economical, simple and efficient cell based ELISA assay for potency estimation of Vedolizumab which has not been reported in any pharmacopoeia. The proposed bioassay method was optimized by investigating Vedolizumab binding to α4β7 integrin which is expressed by HuT78 cells. The validation of this method was done at different parameters including specificity, linearity, range, repeatability, precision, and accuracy. The Vedolizumab binding by ELISA results were found specific for Vedolizumab with linearity (R = 0.99) and precision (%Geometric Coefficient of variance) observed for repeatability and intermediate precision were 3.38% and 2.6% respectively. The relative bias was calculated as 8.68% for repeated performances by different analysts and found in accordance with parameter of accuracy as per various pharmacopoeial guidelines. The developed method is established as robust, effective, and less expensive than high maintenance setup like flow cytometry based assay.
维多珠单抗是一种用于治疗炎症性肠病的人源化单克隆抗体。维多珠单抗与α4β7整合素复合物结合，并抑制其与黏膜地址素细胞黏附分子-1（MAdCAM-1）的结合。为了评估维多珠单抗的结合效力和质量控制检查，使用HuT78细胞进行流式细胞术检测。众所周知，流式细胞仪成本高昂，需要高水平的设备维护以及指定的技术人员来操作。在这方面，本研究的目的是开发并验证一种经济、简单且高效的基于细胞的酶联免疫吸附测定法（ELISA），用于评估维多珠单抗的效价，这在任何药典中均未报道。通过研究维多珠单抗与HuT78细胞表达的α4β7整合素的结合，对所提出的生物测定方法进行了优化。该方法在特异性、线性、范围、重复性、精密度和准确度等不同参数下进行了验证。ELISA结果显示维多珠单抗的结合具有特异性，线性关系良好（R = 0.99），重复性和中间精密度的精密度（几何变异系数%）分别为3.38%和2.6%。不同分析人员重复操作的相对偏差计算为8.68%，根据各种药典指南，该偏差符合准确度参数要求。所开发的方法被确立为稳健、有效，且比基于流式细胞术检测等需要高维护成本的方法更便宜。
