Development of ultra-high sensitivity bioluminescent enzyme immunoassay for prostate-specific antigen (PSA) using firefly luciferase.
A bioluminescent enzyme immunoassay (BLEIA) for prostate-specific antigen (PSA) using biotinylated firefly luciferase-labelled antibody was developed. PSA is an important marker for the diagnosis and management of prostate cancer. Our BLEIA for PSA, based on the two-step sandwich method, had ultra-high sensitivity and a very wide measurable range. The detection limit (mean of nine replicates of the zero standard +2 SD) for PSA was 0.25 pg/mL and the measurable range for PSA was 0.25 pg/mL-100 ng/mL. Generally, PSA in the serum exists on two forms, called free PSA (f-PSA) and complex PSA (c-PSA), which is formed with alpha-antichymotripsin. Thus, the response of the PSA assay to these two forms has to be equimolar in the construction of the assay system. Our BLEIA for PSA also had an equimolar response to them.
Seto
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Biochemical Research Laboratory, Eiken Chemical Co. Ltd., 1381-3 Ohtawara-shi, Tochigi, 324-0036, Japan.
Iba
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Abe
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D000818:Animals; D000911:Antibodies, Monoclonal; D001517:Coleoptera / Q000201:enzymology*; D005260:Female; D006801:Humans; D007124:Immunoenzyme Techniques / Q000379:methods / Q000592:standards*; D018189:Immunomagnetic Separation; D008156:Luciferases; D049449:Luminescence; D017430:Prostate-Specific Antigen / Q000032:analysis* / Q000276:immunology; D012015:Reference Standards; D012680:Sensitivity and Specificity
D000911:Antibodies, Monoclonal; D008156:Luciferases; D017430:Prostate-Specific Antigen