Application of Intra-Assay Calibration Curves to Quantitate Clinical Biomarker Immunoassays
Paul Rhyne,
Associate Director, PCO Bioanalytical Sciences, Biomarkers
Bristol-Myers Squibb
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Abstract: The measurement of biomarkers in clinical samples using immunoassays is typically accomplished using calibration curves generated from known concentrations of standards or calibrators. These standards usually consist of purified protein standards diluted in assay buffer and are run in separate wells. We have developed a different approach using the multiplexing capabilities of the Luminex bead based xMAP technology, where the calibration standards are incorporated into each well along with a clinical sample. This allows for the first time, reference standard information to be generated simultaneously with the measurement of the biomarker in the same well. The benefits of this approach include increased number of clinical samples run on as assay plate, higher replicates of the calibration curve points, and the elimination of buffers dedicated to dilution of reference standards. The principles of this biomarker assay design will be discussed and examples of its application will be shown. Additionally, the benefits and risks of using intra-assay calibration based assays will be discussed.
Presenter Biography: Paul W. Rhyne joined Bristol-Myers Squibb’s Pharmaceutical Research Institute (PRI) in 2005. Paul leads a group of scientists that develop and validate immunoassays that are used to measure biomarkers in support of phase I and early phase II clinical trials. He is also responsible for developing assays for novel biomarkers in support of clinical discovery and other early discovery teams.
Prior to joining Bristol-Myers Squibb, Paul served as an R&D Lab Manager at Upstate Biotechnology in Lake Placid, New York. He was responsible for creating over 40 Luminex based commercial assays (BeadlyteÔ product line) for the detection of cell signaling proteins. Earlier Paul was part of a small start-up biotechnology company (Amplistar Inc.) in North Carolina where he was responsible for developing multiplexed immunoassays that detected anti-cancer antibodies in ovarian cancer patients.
Paul earned his Ph.D. degree in Cellular Immunology at the University of Tennessee Memphis in 1997, focusing on antigen processing and presentation of antigen by B cells. After completing his degree, Paul joined the department of Virology and Molecular Biology as a post-doctoral fellow at St. Jude Children’s Research Hospital in Memphis TN. There he investigated the mechanisms of Epstein-Barr Virus early latency genes in maintaining viral latency and their implicated roles in Burkitt’s lymphoma and other EBV related cancers. Paul has been involved in a broad spectrum of research and has many scientific publications and abstracts that he has authored.
Online Presentation: http://lifescience.planetconnect.com/ppt/LSPrinceton/tuesday/PAUL_RHYNE.ppt
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