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題 名 | Using a Heavy Chain-Loss Hybridoma 26.4.1LL for Studying the Structural Basis of Immunoglobulin Chain Association=利用重鏈缺失的融合瘤細胞26.4.1LL研究免疫球蛋白鏈組合之結構基礎 |
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作 者 | 楊秋英; | 書刊名 | Proceedings of the National Science Council : Part B, Life Science |
卷 期 | 24:3 2000.07[民89.07] |
頁 次 | 頁101-107 |
分類號 | 361.8 |
關鍵詞 | 重鏈缺失; 融合瘤細胞26.4.1LL; 免疫球蛋白鏈; Immunoglobulin; Chain association; Immunoprecipitation; |
語 文 | 英文(English) |
英文摘要 | One of the mechanisms contributing to antibody diversity is created by the association of different heavy and light chains. The combinability of heavy and light chains has been studied previously in two systems: in vitro chain recombination and hybrid hybridoma. Here, a novel in vivo chain combination assay system involving a heavy chain-loss variant, 26.4.1LL, producing two k light chains (LDEX and LMPC) different in size is described. In conjunction with DNA transfection, immunoprecipitation and SDS-PAGE, the structural basis of noncovalent interaction between heavy and light chains can be elucidated systematically by examining the relative association tendency of a heavy chain with two light chains. To demonstrate the usefulness of this system, three stably transfected 26.4.1LL cell lines expressing g2b heavy chains, designated as HDEX, HCHI and HARS, respectively, with structural interrelated variable regions were generated: HDEX differs from HCHI only in framework regions whereas HCHI differs from HARS in complementarity- determining regions. The relative amounts (R values) of LDEX and LMPC associated with the heavy chains HDEX, HCHI and HARS in the assembled immunoglobulin molecules were found to be 1.02, 0.64 and 0.05, respectively, suggesting that the complementarity-determining regions and framework regions contribute equally to the VL-VH interaction. This conclusion is consistent with previous observations based on calculation of the buried area in the VL-VH interface, thus demonstrating the usefulness of this system. |
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