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Development of a Surface Plasmon Resonance‐Based Assay to Detect Antiproliferative Factor in Interstitial Cystitis Patient Urine
Author(s) -
Chavda Burzin,
Ling Jun,
Majernick Thomas,
Planey Sonia
Publication year - 2015
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.29.1_supplement.576.6
Subject(s) - urine , surface plasmon resonance , interstitial cystitis , chemistry , biomarker , mutant , chromatography , cancer research , medicine , urinary system , nanotechnology , biochemistry , materials science , nanoparticle , gene
Interstitial cystitis (IC) is a chronic, painful bladder disease that affects mostly women and is frequently misdiagnosed due to lack of a non‐invasive test to detect the disease. Biomarkers for IC have been described, including antiproliferative factor (APF), a sialoglycopeptide that is detectable in the urine of 95‐97% of IC patients vs. normal controls. Validation of APF as a biomarker for IC has been hindered by the absence of robust assays to detect and measure its concentration in patient urine. In this study, we evaluated the utility of surface plasmon resonance (SPR) to detect and quantitate APF in solution through binding to its cellular receptor, cytoskeleton associated protein 4 (CKAP4). Five his‐tagged CKAP4 mutants were designed according to structural predictions in order to optimize the SPR assay. Each purified mutant was immobilized on a CM5 chip through amine‐coupling, and binding activity to APF and as‐APF (a desialylated APF analogue with full activity) was measured. Four of the CKAP4 mutants demonstrated a quantitative binding response to APF and as‐APF, albeit with altered sensitivities; moreover, as‐APF charged to control patient urine could be detected. Future efforts will focus on assay optimization to detect APF in urine from IC patients. USA MED Research ACQ Activity W81XWH‐13‐1‐0454