Human IgG1 Cγ1 Domain Is Crucial for the Bioactivity of the Engineered Anti-CD20 Antibodies
In this study, we discussed the necessity of human lgG1 Cγ1 domain for recombinant antibody using computeraided homology modeling method and experimental studies. The heavy (VH) and light (VL) chain variable regions of 1-28, a murine IgM-type anti-CD20 mAb, were ligated by linker peptide (Gly4Ser)3 to form the single-chain Fv fragment (scFv). Then, the engineered antibody (LH1-3) was generated by fusing scFv with the entire IgG1 heavy constant regions. The 3-D structure of LH1-3 was modeled using computer-aided homology modeling method and the binding activity of LH1-3 was evaluated theoretically. Compared to the 3-D structure of the Fv fragment of the parent antibody, the conformation of the active pocket of LH1-3 was remained because of the rigid support of Cγ1.Further experimental results of flow cytometry showed that the engineered anti-CD20 antibody possessed specifically binding activity to CD20-expressing target cells. The anti-CD20 antibody fragments could also mediate complement-dependent cytotoxicity (CDC) of human B-lymphoid cell lines. Our study highlights some interests and advantages of a methodology based on the homology modeling and analysis of molecular structural properties.
CD20 engineered antibody binding activity molecular modeling
Shusheng Geng Jiannan Feng Yan Li Xianjiang Kang Yingxun Sun Xin Gu Ying Huang Hong Chang Beifen Shen
College of Life Science, Hebei University, Baoding, China Institute of Basic Medical ciences, Beijin Institute of asic Medical Sciences, Beijing, China Lab of Cellular and Molecular Immunology, Medical Institute of Basic Medical Sciences, Beijing, China Lab of Cellular and Molecular Immunology, Medica College of Life Science, Hebei University, Baoding, China Institute of Basic Medical Sciences, Beijing, China
国际会议
广州
英文
97-101
2008-11-01（万方平台首次上网日期，不代表论文的发表时间）