Preparation of a chemiluminescent imidoester for the non-radioactive labelling of proteins.
A chemiluminescent aryl acridinium ester was synthesized which possesses an imidate ester group capable of reacting with proteins under mild conditions. The compound can be detected at levels as low as 5.2 x 10(-19) mol using commercially available luminometers and can therefore be used to produce high specific activity labelled antibodies for use in immunochemiluminometric assays. The imidate ester compares favourably with a previously reported N-succinimidyl ester in terms of its labelling properties but is easier to synthesize, requiring one less step. The compound was used to label affinity purified to synthesize, requiring one less step. The compound was used to label affinity purified sheep antibodies to human parathyroid hormone to demonstrate its utility in a two-site immunochemiluminometric assay for the measurement of intact parathyroid hormone.
Batmanghelich
S
S
Department of Medical Biochemistry, University of Wales College of Medicine, Health Park, Cardiff, UK.
Brown
R C
RC
Woodhead
J S
JS
Weeks
I
I
Smith
K
K
D000166:Acridines / Q000138:chemical synthesis*; D000906:Antibodies; D006801:Humans; D007096:Imidoesters / Q000138:chemical synthesis*; D007118:Immunoassay / Q000379:methods; D007202:Indicators and Reagents; D008163:Luminescent Measurements; D010281:Parathyroid Hormone / Q000032:analysis*; D010446:Peptide Fragments / Q000032:analysis; D011506:Proteins / Q000032:analysis*; D019379:Teriparatide
D000166:Acridines; D000906:Antibodies; D007096:Imidoesters; D007202:Indicators and Reagents; D010281:Parathyroid Hormone; D010446:Peptide Fragments; D011506:Proteins; D019379:Teriparatide; C075367:4-(2-methoxycarbonimidoylethyl)phenyl 10-methyacridinium-9-carboxylate