Characterisation, crystallisation and preliminary X‐ray diffraction analysis of a Fab fragment of a rat monoclonal antibody with very high affinity for the human muscle acetylcholine receptor

[摘要]

The Fab fragment of a rat monoclonal antibody (no. 192) with very high affinity for the main immunogenic region of the human muscle nicotinic acetylcholine receptor (AChR) has been purified, characterised and crystallised using vapour diffusion techniques. Its Kd for human AChR was determined to be 5×10⁻¹¹ M. Its cross-reactivity pattern suggests that residue α23 of the AChR strongly affects its epitope. Crystals suitable for X-ray analysis, obtained by micro- and macroseeding techniques, belong to the orthorhombic space group C222₁ and they diffract to 2.8 A resolution using synchrotron radiation. The unit cell dimensions are α = 83.4 Å, and and there are two Fab molecules per asymmetric unit.

[学科分类] 生物化学/生物物理

[关键词] Myasthenia gravis;Acetylcholne receptor;Main immunogenic region;Monoclonal antibody;Crystallisation;X-ray crystallography