Detection of luciferase having two kinds of luminescent colour based on optical filter procedure: application to an enzyme immunoassay

This paper reports on our study using several optical filters known to be e fficient in separating compounds having various levels of maximum luminesce nce, to separate information from three kinds of Luciola lateralis lucifera se with a maximum luminescence of 559 nm, 604 nm and 607 nm. Simultaneous l uminescence of Luciola lateralis luciferase was determined by measuring the luminescence through a band pass filter or sharp cut filter (BPB50, 53, 58 , No.58, SC58, 60, 62, 64). It was possible to determine luciferase with a maximum luminescence lambda(max) of 559 nm (yellow-green) utilizing the ban d pass filter (BPB 50), described here, Meanwhile, luciferase with a lambda (max) of 607 nm (red) could be determined by calculations based on the biol uminescent intensity through the band pass filter and sharp cut filter (SC5 8). In addition, we also applied a simultaneous bioluminescent enzyme immun oassay of pepsinogen I (PGI) and pepsinogen II (PGII) in which two kinds of biotinylated luciferase (Luciola lateralis) labelled as an enzyme producin g yellow-green light (lambda(max) = 559 nm) and red light (lambda(max) = 60 7 nm) were used. In the proposed method, PGI and PGII in serum were simulta neously captured in a sandwich-type immune reaction between anti-PGI and an ti-PGII monoclonal antibody-coated magnetic particles, and streptavidin-bio tinylated luciferase biotinylated anti-PGI and anti-PGII monoclonal antibod ies triplexes, respectively. The result was a calibration range for PGI of 2-200 ng/mL, and for PGII of 1-100 ng/mL. In conclusion, the correlation of PG values in serum between the proposed method (simultaneous assay) and an individual specific bioluminescent immunoassay (specific assay) were satis factory. Copyright (C) 2000 John Wiley & Sons, Ltd.