Lung Cancer-Associated Keratin 19 Fragments: Development and Biochemical Characterisation of the New Serum Assay Enzymun-Test® Cyfra 21–1

Author:

Bodenmüller H.1,Ofenloch-Hähnle B.1,Lane E.B.2,Dessauer A.1,Böttger V.2,Donié F.1

Affiliation:

1. Boehringer Mannheim GmbH, Research Center Tutzing, 82327 Tutzing - Germany

2. CRC Laboratories, Dept. of Anatomy and Physiology, University of Dundee, Scotland - UK

Abstract

From a panel of 4 murine monoclonal antibodies directed against keratin 19 various antibody combinations were evaluated in solid-phase enzyme-linked sandwich immunoassays for detection of soluble keratin 19 fragments in patient sera. One of these antibody combinations, comprised of the monoclonal antibodies Ks 19.1 and BM 19.21, was selected for further development to a routine test (Enzymun-Test® CYFRA 21–1) because of its high diagnostic sensitivity and specificity for non-small cell lung carcinoma (NSCLC). Both antibodies are specific for keratin 19, no reactivity could be observed with cytokeratin 8 or 18. The epitopes of the two antibodies were determined to be within helix 2B of the rod romain. The epitope sequences lie within the sequence 311–335 for the catcher antibody Ks 19.1 and 346–367 for the detector antibody BM 19.21. These sequences are unique, as could be confirmed from sequence databases. The standard material for the assay was prepared from a cytoskeleton fraction of cultivated MCF-7 cells. Subsequent digestion of this fraction with chymotrypsin yielded a soluble and stable standard material. Both the standard material and the serum analyte appeared as oligomers when analysed on gel chromatography: the serum analyte appeared exclusively at a Mr of 100 ± 10 kD, whereas the standard material eluted in fractions corresponding to 100 ±10 kD and 450 kD. Due to the precise definition of the antigen and the localisation of the antibody binding sequences, Enzymun-Test® CYFRA 21–1 is one of the best characterised tumor markers so far.

Publisher

SAGE Publications

Subject

Cancer Research,Clinical Biochemistry,Oncology,Pathology and Forensic Medicine

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