Automated Time-Resolved Immunofluorometric Assay for Progastrin-Releasing Peptide

Small cell lung cancer accounts for approximately 20% of new cases of lung cancer, and advanced disease is prevalent at the time of diagnosis. Neuron-specific enolase (NSE) has been the primary tumor marker in small cell lung cancer but it has relatively low sensitivity in early-stage disease. Proga...

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Bibliographic Details
Published in:Clinical chemistry (Baltimore, Md.) Md.), 2008-05, Vol.54 (5), p.919-922
Main Authors: Nordlund, Marianne S, Warren, David J, Nustad, Kjell, Bjerner, Johan, Paus, Elisabeth
Format: Article
Language:English
Subjects:
Amino acids
Analytical, structural and metabolic biochemistry
Animals
Antibodies, Monoclonal - isolation & purification
Autoanalysis
Biological and medical sciences
Biomarkers, Tumor - blood
Carcinoma, Small Cell - diagnosis
E coli
Enzyme-Linked Immunosorbent Assay
Female
Fluoroimmunoassay
Fundamental and applied biological sciences. Psychology
Gastrin-Releasing Peptide - blood
Gastrin-Releasing Peptide - immunology
Humans
Immunization
Investigative techniques, diagnostic techniques (general aspects)
Lung cancer
Lung Neoplasms - diagnosis
Medical sciences
Mice
Mice, Inbred BALB C
Monoclonal antibodies
Peptides
Phosphopyruvate Hydratase - blood
Polyethylene glycol
Protein Precursors - blood
Protein Precursors - immunology
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Summary:Small cell lung cancer accounts for approximately 20% of new cases of lung cancer, and advanced disease is prevalent at the time of diagnosis. Neuron-specific enolase (NSE) has been the primary tumor marker in small cell lung cancer but it has relatively low sensitivity in early-stage disease. Progastrin-releasing peptide (proGRP) is a promising alternative or complementary marker for NSE. We have previously described a time-resolved immunofluorometric assay (TR-IFMA) for proGRP that lacked the necessary sensitivity and robustness for use in the routine clinical laboratory. Herein we describe the development of an improved assay using a novel monoclonal antibody pair. Mice were immunized with different conjugated proGRP peptides, including residues 31-98, 1-98, and preproGRP(-23-125). Pair combinations of the resulting monoclonal antibodies (mAb) were tested. The improved TR-IFMA was compared with the only other available proGRP assay, the proGRP ELISA (IBL). A panel of 12 high-affinity mAbs was produced. The best assay combination was between our original E146 mAb as solid-phase antibody and the new mAb M16 as tracer. The new TR-IFMA had a linear dose-response curve, a wide dynamic range (13-13 500 ng/L), and a limit of detection of 2.8 ng/L. Total CV was
ISSN:0009-9147
1530-8561
DOI:10.1373/clinchem.2007.101436