Department of Medical Biotechnology and Nanotechnology, Faculty of Medicine, Zanjan University of Medical Sciences, Zanjan, Iran.
Department of Clinical Biochemistry, Faculty of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
J Cell Biochem. 2019 Jan;120(1):940-950. doi: 10.1002/jcb.27457. Epub 2018 Aug 30.
CD20 molecule, a phosphoprotein with 297 amino acids and four transmembrane domains, is a member of MS4A protein family. Anti-CD20 antibodies such as ofatumumab, which have been developed for cancer treatment and has demonstrated efficacy in relapsed/refractory chronic lymphocytic leukemia, are among the most successful therapies to date. Rational engineering methods can be applied with reasonable success to improve functional characteristics of antibodies. Considering the importance of this issue, we have used in silico modeling approach for the improvement of ofatumumab monoclonal antibody. Four mutated variants of ofatumumab were developed and expressed in Chinese hamster ovary (CHO) cells along with the unmodified antibody. Analysis of affinity of the purified antibodies with CD20 showed significant improvement in antigen-binding characteristics of one of the variants compared with the control antibody. This study represents the first step toward development of the second generation ofatumumab antibody with improved affinity.
CD20 分子是一种由 297 个氨基酸和四个跨膜结构域组成的磷酸化蛋白，属于 MS4A 蛋白家族。抗 CD20 抗体，如奥法木单抗，已被开发用于癌症治疗，并已证明在复发性/难治性慢性淋巴细胞白血病中有疗效，是迄今为止最成功的治疗方法之一。合理的工程方法可以合理地应用于改善抗体的功能特性。鉴于这个问题的重要性，我们已经使用计算机模拟方法来改进奥法木单抗单克隆抗体。我们设计并在 CHO 细胞中表达了奥法木单抗的四个突变体，以及未修饰的抗体。对纯化抗体与 CD20 的亲和力进行分析，结果显示与对照抗体相比，其中一个变体的抗原结合特性有了显著提高。这项研究代表了开发具有改善亲和力的第二代奥法木单抗抗体的第一步。
