Tyler Yin (Presenter)
University of Louisville
Bio: Dr. Tyler Yin is a second year clinical chemistry fellow at the University of Louisville. His current research interests are in developing assays for biomarker detection using mass spectrometry. Before joining the clinical chemistry program, his main research focus was in the area of enzyme engineering and discovery through which he has made important contributions.
Authorship: (1)Delu (Tyler) Yin, (1)James Hribar, (1)Michael Merchant, (1)(2)Roland Valdes Jr.
(1) University of Louisville, (2) PGXL Technologies
| Short Abstract Obstructive sleep apnea (OSA) is a disorder where a person is deprived of oxygen that may cause additional health problems if left untreated. Children with OSA are thought to be underdiagnosed due to the lack of sleep centers around the country and noncompliance with take-home testing. Urocortin-3, (UCN3) has been previously identified as a urine biomarker for OSA in children. Here, we utilized multiple reaction monitoring to identify a peptide fragment that gave 3 transitions with high signal intensity. We find that the linearity and sensitivity are sufficient to make a differentiation between children with and without OSA based on previous findings. We also developed two methods in parallel that deplete abundant proteins and concentrate UCN3 in both urine and plasma spiked samples. Both methods yield highly purified UCN3 that are scalable for high throughput assays. |
Long Abstract
Obstructive sleep apnea (OSA) is a disorder where a person is deprived of oxygen that may cause additional health problems if left untreated. Children with OSA are thought to be underdiagnosed due to the lack of sleep centers around the country and noncompliance with take-home testing. Urocortin-3, (UCN3) has been previously identified as a urine biomarker for OSA in children. Here, we utilized multiple reaction monitoring to identify a peptide fragment that gave 3 transitions with high signal intensity. We find that the linearity and sensitivity are sufficient to make a differentiation between children with and without OSA based on previous findings. We also developed two methods in parallel that deplete abundant proteins and concentrate UCN3 in both urine and plasma spiked samples. For plasma, we find that UCN3 is retained in the supernatant with 80% acetonitrile, while depleting up to 97% of other proteins. Application of this method to urine yielded an increased concentration of urobilinogen, which interferes with downstream applications. The high isoelectric point (pI=11.5) allowed us to purify the peptide using a weak cation exchanger. We find that UCN3 has a strong affinity towards carboxymethyl cellulose resins which requires up to 1.3 M NaCl with 10% acetonitrile to elute. Together, both methods yield highly purified UCN3 that are scalable for high throughput assays.
References & Acknowledgements:
We thank the PGXL foundation for their financial support in this project.
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IP Royalty: no
| Planning to mention or discuss specific products or technology of the company(ies) listed above: | no |