与免疫原性半抗原和交叉反应性半抗原复合的抗4-羟基-3-硝基苯乙酸单克隆抗体Fab片段的结晶及初步X射线衍射研究

Crystallization and preliminary X-ray diffraction studies of an anti-4-hydroxy-3-nitrophenylacetic acid monoclonal antibody Fab fragment complexed with immunizing and heteroclitic haptens.

作者信息

Yuhasz S C, Ysern X, Strand M, Amzel L M

机构信息

Department of Biophysics, Johns Hopkins University, School of Medicine, Baltimore, MD 21205.

出版信息

J Mol Biol. 1989 Sep 20;209(2):319-21. doi: 10.1016/0022-2836(89)90280-5.

DOI:10.1016/0022-2836(89)90280-5

PMID:2585486

Abstract

The Fab fragment of the anti-4-hydroxy-3-nitrophenylacetic acid monoclonal antibody, 88C6/12 has been crystallized in the presence of the eliciting hapten, 4-hydroxy-3-nitrophenacetyl-epsilon-aminocaproic acid (NP-aminocap) and the heteroclitic iodinated analog, 4-hydroxy-3-iodo-5-nitrophenylacetyl-epsilon-aminocaproic acid (NIP-aminocap). Crystals obtained by precipitation with 32% (w/v) polyethylene glycol 3400 in the presence of 40 to 400 microM of either NP-aminocap or NIP-aminocap, belong to the orthorhombic space group P2(1)2(1)2(1) with cell dimensions a = 81.2 A, b = 86.9 A, c = 131.1 A. The cell volume suggests the presence of two molecules of the complex per asymmetric unit. Analysis of the Patterson function indicates that these two molecules are related by a local 2-fold axis parallel to the crystallographic b axis located at x = 0.218 and z = 0.25.

摘要

抗4-羟基-3-硝基苯乙酸单克隆抗体88C6/12的Fab片段在引发半抗原4-羟基-3-硝基苯乙酰基-ε-氨基己酸（NP-氨基己酸）和异向性碘化类似物4-羟基-3-碘-5-硝基苯乙酰基-ε-氨基己酸（NIP-氨基己酸）存在的情况下结晶。在40至400微摩尔的NP-氨基己酸或NIP-氨基己酸存在下，通过用32%（w/v）聚乙二醇3400沉淀获得的晶体属于正交晶系空间群P2(1)2(1)2(1)，晶胞尺寸a = 81.2 Å，b = 86.9 Å，c = 131.1 Å。晶胞体积表明每个不对称单元存在两个复合物分子。帕特森函数分析表明，这两个分子通过平行于位于x = 0.218和z = 0.25的结晶学b轴的局部2重轴相关。