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.
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 X 10(-11) M. Its cross-reactivity pattern suggests that residue alpha23 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 C222(1) and they diffract to 2.8 A resolution using synchrotron radiation. The unit cell dimensions are alpha=83.4 A, b=110.0 A and c=212.2 A and there are two Fab molecules per asymmetric unit.
Kontou
M
M
Department of Biochemistry, Hellenic Pasteur Institute, Athens, Greece.
Vatzaki
E H
EH
Kokla
A
A
Acharya
K R
KR
Oikonomakos
N G
NG
Tzartos
S J
SJ
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry; D000915:Antibody Affinity; D003429:Cross Reactions; D003460:Crystallization; D018360:Crystallography, X-Ray; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000737:chemistry* / Q000302:isolation & purification; D009132:Muscles / Q000737:chemistry / Q000276:immunology; D051381:Rats; D011950:Receptors, Cholinergic / Q000276:immunology*
D000911:Antibodies, Monoclonal; D007140:Immunoglobulin Fab Fragments; D011950:Receptors, Cholinergic