Validation of APF as a Urinary Biomarker for Interstitial Cystitis
Abstract
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.
Document Details
- Document Type
- Technical Report
- Publication Date
- Oct 01, 2015
- Accession Number
- ADA626134
Entities
People
- Sonia L. Planey
Organizations
- The Commonwealth Medical College