Ting Yi Tian, Temme Sebastian, Koch Norbert, McLellan Alexander D
Department of Microbiology and Immunology, University of Otago, Dunedin, New Zealand.
Hybridoma (Larchmt). 2009 Dec;28(6):423-9. doi: 10.1089/hyb.2009.0050.
We report the generation of a monoclonal antibody (MAb) that reacts to the N-terminus of the denatured HLA-DRalpha chain. The 1C4.6 MAb was raised against a peptide corresponding to amino acid residues 10 to 32 of a highly conserved region within the alpha1 domain of HLA-DR. This region partially overlaps with the epitope recognized by the conformationally dependent L243 MAb. In Western blot analysis, MAb 1C4.6 reacted with denatured HLA-DRalpha chains, but failed to bind the HLA-DRbeta chain expressed individually by transfectant cells, confirming that it recognizes an epitope on the alpha-chain of HLA-DR. In addition, this antibody was found to be isotype specific to HLA-DRalpha, as it did not cross-react to HLA class II proteins HLA-DP and-HLA-DQ. The 1C4.6 MAb is a valuable addition to existing reagents used to probe the structure and function of MHC class II molecules. This anti-HLA-DRalpha1 domain MAb may prove valuable for studies of HLA class II heterodimer assembly, structure, and function, as well as for studies into the release of soluble MHC class II.
我们报告了一种单克隆抗体（MAb）的产生，该抗体与变性的HLA-DRα链的N端发生反应。1C4.6单克隆抗体是针对与HLA-DRα1结构域内高度保守区域的氨基酸残基10至32相对应的肽产生的。该区域与构象依赖性L243单克隆抗体识别的表位部分重叠。在蛋白质印迹分析中，1C4.6单克隆抗体与变性的HLA-DRα链发生反应，但未能结合转染细胞单独表达的HLA-DRβ链，证实它识别HLA-DRα链上的一个表位。此外，发现该抗体对HLA-DRα具有同种型特异性，因为它与HLA II类蛋白HLA-DP和-HLA-DQ不发生交叉反应。1C4.6单克隆抗体是用于探测MHC II类分子结构和功能的现有试剂的宝贵补充。这种抗HLA-DRα1结构域单克隆抗体可能对HLA II类异二聚体组装、结构和功能的研究以及可溶性MHC II类释放的研究具有重要价值。
