Simultaneous assay of pepsinogen I and pepsinogen II in serum by bioluminescent enzyme immunoassay using two kinds of Luciola lateralis luciferase

Ohkuma, H Abe, K Kosaka, Y Maeda, M

H. Ohkuma et al., Simultaneous assay of pepsinogen I and pepsinogen II in serum by bioluminescent enzyme immunoassay using two kinds of Luciola lateralis luciferase, ANALYT CHIM, 395(3), 1999, pp. 265-272

17

INGLESE

Article

Spectroscopy /Instrumentation/Analytical Sciences

ANALYTICA CHIMICA ACTA

0003-2670 → ACNP

395

3

1999

265 - 272

ISI

0003-2670(19990823)395:3<265:SAOPIA>2.0.ZU;2-0

We developed a simultaneous bioluminescent enzyme immunoassay of serum peps inogen I (PG I) and pepsinogen II (PG II) in which two kinds of biotinylate d luciferase (Luciola lateralis) as labeled enzyme producing yellow-green l ight (lambda(max)=559 nm) and red light (lambda(max)=607 nm) were used. In the proposed method, PG I and PG II in serum were simultaneously captured i n a sandwich type immunereaction between anti-PG I and anti-PG II monoclona l antibody-coated magnetic particles, and streptavidin biotinylated lucifer ase biotinylated anti-PC I and anti-PG II monoclonal antibodies tripler, re spectively. After bound/free (B/F)separation by washing, bound enzyme activ ities of luciferase were measured twice as the bioluminescent intensity in the presence of luciferin, ATP, Mg2+ and molecular oxygen. The value of PG I was obtained by calculation based on the bioluminescent intensity at lamb da(max)=607 nm. The value of PG II (lambda(max)=559 nm) was obtained by the method using a band pass filter. The calibration range of PG I was from 2 to 200 ng ml(-1) and that of PG II was from 1 to 100 ng ml(-1). The coeffic ients of variation for PG I and PG II determination were from 3.4% to 10.2% and from 4.0% to 8.6%, respectively. The correlation of PG I values in ser um between the proposed method and a PG I specific bioluminescent immunoass ay (single assay), and PG II values in serum between the proposed method an d a PG II specific bioluminescent immunoassay were satisfactory. The regres sions were y(proposed method)=0.9952x(single assay for PY I)+0.841, r=0.983 7 and y(proposed method)=1.0545x(single assay for PY II)+1.4693, r=0.9738, respectively. The proposed method is concise, accurate and rapid for the de termination of PG I and pc II in serum, and is useful for clinical applicat ion. (C) 1999 Published by Elsevier Science B.V. All rights reserved.