Abstract

Article info 2017 5 (2)    005  (02)   pp.  66 ~ 69
Title Expression, purification, crystallization, and preliminary crystallographic analysis of the Fab fragment of golimumab, a therapeutic antibody against TNFα
Authors Jee Un Lee, Ji Young Son, Woori Shin, Ki-Young Yoo, Heejin Lim, Ju Yeon Lee, Hyun Tae Lee and Yong-Seok Heo*
Institutions Department of Chemistry, Konkuk University, 120 Neungdong-ro, Gwangjin-gu, Seoul 05029, Republic of Korea. *Correspondence: ysheo@konkuk.ac.kr
Abstract TNFα is an inflammatory cytokine required for immune responses, playing a crucial role in the pathogenesis of inflammatory autoimmune diseases via interactions with its cognate receptors, TNFR1 and TNFR2. Therapeutic antibodies targeting TNFα, including infliximab, adalimumab, certolizumab pegol, and golimumab, are clinically used for the treatment of the inflammatory diseases such as rheumatoid arthritis, psoriatic arthritis, Crohn’s disease, and inflammatory bowel disease. Here, the Fab fragment of golimumab was overexpressed in the periplasmic region of Escherichia coli and purified by affinity and gel-filtration chromatography. The purified protein was crystallized by hanging-drop vapor diffusion in the presence of 100 mM Tris, pH 8.5, 0.1 M ammonium phosphate, and 12% (w/v) PEG 6,000 at 20°C. The crystal diffracted X-ray to 2.60 Å resolution and belonged to the orthogonal space group P2 1 2 1 2, with unit cell parameters a = 72.74, b = 130.29, and c = 93.35 Å. An asymmetric unit of the crystal contains two molecules of the Fab fragment of golimumab with a V M of 2.14 Å 3 Da -1 and a solvent content of 42.48%.