Journal
BRAIN RESEARCH
Volume 895, Issue 1-2, Pages 277-282Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/S0006-8993(01)02010-8
Keywords
human brain; acetylcholinesterase; polyclonal antibody; combinatorial chemical synthesis; enzyme-linked immunosorbent assay; epitope; synthetic peptide; cross-immunoreactivity
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Antigenic decapeptides of human brain acetylcholinesterase (AChE) were investigated for immunoreactivity with the rabbit anti Torpediniformes narcine timlei AChE polyclonal antibody (anti-narcine AChE polyclonal antibody). The decapeptides were synthesized using the multipin combinatorial chemical synthesis technique and biotinylated at N-terminals. Rabbit anti-narcine AChE polyclonal antibodies were purified by Protein A-Sepharose CL 4B column chromatography. Enzyme-linked immunosorbent assay (ELISA) was used for the assay of the reaction between the antigen and the antibody. Seven of 11 antigenic synthetic decapeptides of human brain AChE showed obvious immunoreactivity with the rabbit anti-narcine AChE polyclonal antibodies. The similarity of the AChE sequences of humans and Torpedo species were compared thereby with the epitopes indicated. The results indicate that the epitopes of human brain AChE and Torpedo AChEs have been highly conserved during evolution. In view of this, no N-glycosylation site was found in the antigenic decapeptides tested, they all belong to oligopeptide epitopes. (C) 2001 Elsevier Science B.V. All rights reserved.
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