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首页> 外文期刊>Acta crystallographica.Section D. Biological crystallography >Combined use of AFM and X-ray diffraction to analyze crystals of an engineered, domain-deleted antibody
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Combined use of AFM and X-ray diffraction to analyze crystals of an engineered, domain-deleted antibody

机译：利用AFM和x射线衍射相结合分析晶体的一个工程,domain-deleted抗体

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摘要

A genetically engineered humanized C(H)2-domain-deleted monoclonal antibody lacking any interchain-hinge disulfide bonds has been crystallized in the presence of detergent in a form suitable for X-ray diffraction analysis. The crystals were grown from 4 M formate along with Triton X-100 and had P2(1)2(1)2 space-group symmetry, with unit-cell parameters a = 83, b = 224, c = 167 angstrom. The crystals diffract to beyond 2.8 angstrom resolution. A disordered crystal form of larger size and more attractive habit was also grown from 4 M formate, but in the presence of the Anapoe series of detergents. Preliminary X-ray data, in conjunction with atomic force microscopy images, are consistent with asymmetric units consisting of two intact antibodies forming a circular dimeric ring. The crystallizing unit, which must contain a twofold axis, is a toroidal assembly of four antibodies (two dimeric rings). Competition between dimers and tetramers to enter the lattice, along with a unique kind of planar defect of packing, may be responsible for the unusually high defect density and the disorder of the X-ray diffraction pattern exhibited by the second crystal form. An approach to crystallizing proteins showing phase separation, particularly intact antibodies, that uses a preliminary detergent test set is described.
机译：一种转基因人性化C (H) 2-domain-deleted单克隆抗体缺乏任何interchain-hinge二硫键结晶在洗涤剂的存在形成适合x射线衍射分析。甲酸以及晶体从4米特里同x - 100和P2(1) 2(1) 2空间群对称,晶胞参数= 83,b =224年,c = 167埃。超出2.8埃分辨率。水晶的更大的规模和形式更有吸引力甲酸的习惯也是从4米,但在Anapoe存在一系列的洗涤剂。初步的x射线数据,结合原子力显微镜图像,是一致的与不对称单位组成的两个完好无损抗体形成一个环状二聚的戒指。结晶单元,它必须包含一个双重的四轴,是一个环形组装抗体(两个二聚的环)。和四聚体进入晶格,以及独特的包装,平面缺陷的可能负责异常高的缺陷密度和x射线衍射模式的障碍第二晶体的形式展出。结晶蛋白质展示阶段分离,特别完整的抗体,使用一个测试集是初步的洗衣粉描述。

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