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Validation of APF as a Urinary Biomarker for Interstitial Cystitis

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Annual rept. 30 Sep 2014-29 Sep 2015

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The purpose of this study is to develop and characterize a surface plasmon resonance SPR-based assay that can specifically detect binding of APF to its cellular receptor, cytoskeleton associated protein 4 CKAP4, immobilized on a sensor chip surface and to test the ability of this SPR-based assay to discriminate and measure the concentration of APF in urine from well-defined IC patients vs. age-matched, asymptomatic controls. In the second year of study, we have nearly completed our patient recruitment and urine sample collection. Testing of these samples by the cellular proliferation assay is underway at the University of Maryland. Further, considerable progress has been made toward the development, characterization, and eventual testing of clinical samples by the SPR assay. The focus in year two has been on development of the SPR assay using CKAP4 as a biosensor to detect APF. Our results demonstrate that we have successfully optimized rCKAP4 activity and immobilization with sufficient binding efficiency to detect and quantitate APF in a purified system. Importantly, we have determined that CKAP4127-360 improves binding to APF with a normal proportional response to increasing dose of APF and the maximum binding response Rmax. A parallel approach using an APF mAb as a biosensor was pursued to enhance sensitivity of the assay it offers an alternate strategy to specifically measure APF in urine with the goal of developing a non-invasive, point-of-care diagnostic test for IC.

Subject Categories:

  • Biochemistry
  • Anatomy and Physiology
  • Medicine and Medical Research

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