单克隆抗肿瘤抗体BR96及其与Lewis Y决定簇复合物的结晶与X射线初步分析

Crystallization and preliminary X-ray analysis of the monoclonal anti-tumor antibody BR96 and its complex with the Lewis Y determinant.

作者信息

Chang C Y, Jeffrey P D, Bajorath J, Hellström I, Hellström K E, Sheriff S

机构信息

Bristol-Myers Squibb Pharmaceutical Research Institute, Princeton, NJ 08543-4000.

出版信息

J Mol Biol. 1994 Jan 7;235(1):372-6. doi: 10.1016/s0022-2836(05)80044-0.

DOI:10.1016/s0022-2836(05)80044-0

PMID:8289260

Abstract

The monoclonal anti-tumor antibody BR96 binds a tetrasaccharide, Lewis y (Le(y)), in vitro and recognizes a Le(y)-bearing or Le(y)-related tumor-associated antigen in vivo. The Fab of the murine monoclonal antibody, mBR96 (IgG3, kappa), and the Fab' of its human chimera, cBR96 (IgG1, kappa), and their complexes with Le(y) have been screened for crystallization conditions. Crystals suitable for X-ray diffraction have been obtained for uncomplexed cBR96 Fab', cBR96 Fab' in complex with Le(y) and mBR96 Fab in complex with Le(y). The symmetry of the cBR96 Fab' crystals is consistent with space group P2(1)2(1)2, a = 61.1 A; b = 174.3 A; c = 45.6 A; the symmetry of the cBR96 Fab'-Le(y) complex crystals with space group P4(3)2(1)2 (or its enantiomorph), a = b = 82.2 A; c = 167.1 A and the symmetry of the mBR96 Fab-Le(y) complex crystals with space group P2(1)2(1)2(1), a = 69.4 A; b = 84.9 A; c = 86.8 A.

摘要

单克隆抗肿瘤抗体BR96在体外与四糖Lewis y（Le(y)）结合，并在体内识别携带Le(y)或与Le(y)相关的肿瘤相关抗原。已对鼠单克隆抗体mBR96（IgG3，κ）的Fab片段、其人源化嵌合体cBR96（IgG1，κ）的Fab'片段及其与Le(y)的复合物进行了结晶条件筛选。已获得适合X射线衍射的未复合cBR96 Fab'、与Le(y)复合的cBR96 Fab'以及与Le(y)复合的mBR96 Fab的晶体。cBR96 Fab'晶体的对称性与空间群P2(1)2(1)2一致，a = 61.1 Å；b = 174.3 Å；c = 45.6 Å；与Le(y)复合的cBR96 Fab'-Le(y)晶体的对称性与空间群P4(3)2(1)2（或其对映体）一致，a = b = 82.2 Å；c = 167.1 Å；与Le(y)复合的mBR96 Fab-Le(y)晶体的对称性与空间群P2(1)2(1)2(1)一致，a = 69.4 Å；b = 84.9 Å；c = 86.8 Å。