Complement Factor H as a Marker for Detection of Bladder Cancer

• Published in: Clinical chemistry (Baltimore, Md.) Md.), 2005-05, Vol.51 (5), p.856-863
• Main Authors: Cheng, Zhu-Zhu, Corey, Michael J, Parepalo, Maria, Majno, Sandra, Hellwage, Jens, Zipfel, Peter F, Kinders, Robert J, Raitanen, Mika, Meri, Seppo, Jokiranta, T. Sakari
• Format: Article
• Language: English
• Subjects: Analytical, structural and metabolic biochemistry; Antibodies, Monoclonal - metabolism; Antigens, Neoplasm - urine; Binding Sites, Antibody; Biological and medical sciences; Biomarkers, Tumor - urine; Bladder cancer; Blood Proteins - metabolism; Blood Proteins - urine; Carcinoma, Transitional Cell - diagnosis; Complement C3b Inactivator Proteins; Complement Factor H - immunology; Complement Factor H - metabolism; Complement Factor H - urine; Fundamental and applied biological sciences. Psychology; Humans; Investigative techniques, diagnostic techniques (general aspects); Ligands; Male; Medical sciences; Protein Binding; Protein Structure, Tertiary; Surface Plasmon Resonance; Urinary Bladder Neoplasms - diagnosis; Urine
• ISSN: 0009-9147; 1530-8561
• DOI: 10.1373/clinchem.2004.042192

The BTA TRAK and BTA stat tests for bladder cancer use monoclonal antibodies (mAbs) X13.2 and X52.1 to detect factor H (FH)-related material in urine. The exact ligands remain unknown. Western blot analyses of purified FH, recombinant factor H-related protein 1 (FHR-1), and serum and urine samples were used to identify the ligands of X13.2 and X52.1. Recombinant FH constructs were used to identify the target sites of X13.2 and X52.1. To analyze whether natural ligands of FH could compete with its recognition by the capture mAb X52.1, we used surface plasmon resonance analysis. The role of the ligands of X52.1 in the BTA TRAK assay was tested with use of purified proteins and FH-depleted samples. X13.2 bound to domain 3 of FH and FH-like protein 1, whereas X52.1 bound to domain 18 of FH and to FHR-1. Using specific FH depletion from a bladder cancer patient's urine and purified FH, we demonstrated that FH is the ligand recognized by the BTA TRAK test. By contrast, FHR-1 in urine reduced the FH-dependent test signal. FH is a tumor marker for bladder cancer. To reveal the presence of bladder cancer, the BTA TRAK assay detects FH, whereas FHR-1 is able to partly inhibit this detection. This indicates a special mechanism for a diagnostic immunoassay based on the combined effect of simultaneous positive and negative signals in a single sample.