Anti-epitope antibody, a novel site-directed antibody against human acetylcholinesterase.
OBJECTIVE
To construct synthetic antigens using the epitope of human brain acetylcholinesterase (hbAChE) for induction and detection of the specific antibody against the epitope, and to analyse the immunogenicity of the antibody.


METHODS
The epitope (RTVLVSMNYR, amino acids 143-152) of hbAChE was chemically synthesized, coupled with the carrier protein keyhole limpet hemocyanin (KLH) to construct an artificial immunogen (KLH-epitope), and injected into rabbits to raise antibody. The epitope conjugated with bovine serum albumin (BSA) was used as the detection antigen. The specificity of the antibody was tested by enzyme-linked immunosorbent assay (ELISA) and Western blotting. The immunoreaction between the anti-recombinant human butyrylcholinesterase (rhBChE) polyclonal antibody and the biotinylated-epitope was examined by indirect ELISA.


RESULTS
The erythrocyte AChE, the hbAChE, rhBChE and the BSA-epitope all immunoreacted with the anti-epitope antibody against the epitope (143-152) of hbAChE, whereas the torpedo AChE did not.


CONCLUSIONS
The hbAChE, the human erythrocyte AChE and hBChE share the conservative antigenic epitope RTVLVSMNYR, hence they can all immunoreact with the anti-epitope antibody. Since the epitope of hbAChE is less similar with the aligned amino acid sequences of AChE of Torpedo californica or Torpedo marmorata, there is not any immunoreactivity between them. The R, M, and N residues in the epitope seem to be necessary radicals for the conservation of antigenicity.
Zhang
Xing-mei
XM
Institute of Pharmacology and Toxicology, Academy of Military Medical Sciences, Beijing 100850, China.
Liu
Gang
G
Sun
Man-ji
MJ
D000110:Acetylcholinesterase / Q000737:chemistry / Q000276:immunology*; D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000737:chemistry; D000918:Antibody Specificity / Q000276:immunology*; D001921:Brain / Q000201:enzymology*; D000939:Epitopes / Q000737:chemistry / Q000276:immunology*; D006801:Humans; D008297:Male; D008969:Molecular Sequence Data; D011817:Rabbits; D017386:Sequence Homology, Amino Acid; D014101:Torpedo / Q000276:immunology
D000906:Antibodies; D000939:Epitopes; D000110:Acetylcholinesterase