Aberrations in protein expression can often lead to disease. Examples include protein mutations, deficiency, over expression, misfolding and aggregation. Amyloids are formed by misfolding of extracellular proteins and deposition of amyloid fibrils in soft tissues causing organ dysfunction. Eliminating the production of the specific proteins indicates a change in the condition. Therefore, unequivocal identification of the amyloid protein is critical to understanding the disease.
The investigation usually starts with Congo red staining followed by protein typing with immunohistochemistry. However, there are significant challenges associated with antibody based subtyping. First, most antibodies are produced against wild-type proteins while amyloid proteins often have mutation, truncation, and conformational changes causing reduced reactivity. Second, antibodies for certain proteins may not be readily available, making it difficult to identify rare proteins.
Protein subtyping by LC/MS offers unique advantages. It is not limited by antibody availability and is able to identify the entire proteome at a single analysis with high sensitivity and specificity.
Dr. Wang's lab at the Cleveland Clinic recently described a novel method for subtyping amyloid proteins using nanoliquid chromatography coupled with a benchtop LC-MS/MS that combines high-performance quadrupole precursor selection with high resolution, accurate-mass detection. This clinical research method offers high sensitivity and specificity for identifying amyloid proteins and will be the focus of the discussion.
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Sihe Wang, PhD, DABCC, FACB
Dr. Sihe Wang is Section Head and Medical Director of Clinical Biochemistry and Director of Clinical Biochemistry Fellowship Training Program, Cleveland Clinic, Cleveland, OH. He also chairs the clinical chemistry integration effort for the Cleveland Clinic Health System, which includes one Florida hospital, eight community hospitals and 18 family health centers in Northeast Ohio. Additionally, he is Clinical Chemistry Professor, Cleveland State University. Prior to his current position, Dr. Wang was Assistant Professor at Northwestern University; Director, Clinical Chemistry Laboratory and Referred Testing Laboratory, Children's Memorial Hospital, Chicago, Illinois. Dr. Wang is a diplomate of the American Board of Clinical Chemistry (DABCC) and a fellow of the National Academy of Clinical Biochemistry (FACB).
Dr. Wang is a member of several professional organizations, including the American Society for Mass Spectrometry and the American Association for Clinical Chemistry (AACC). He served as chair of AACC Northeast Ohio Section in 2008 and 2009 and the president of North American Chinese Clinical Chemistry Association (NACCCA) 2008-2009. Currently he serves as the historian for NACCCA, the treasurer for the Pediatric and Maternal Fetal Division of AACC, the delegate for AACC Northeast Ohio section, commissioner for The Commission on Accreditation in Clinical Chemistry (ComACC), and a member of AACC's Strategies Online Editorial Advisory Board. The AACC presented him with the 2005, 2008, and 2010 Clinical Chemist Recognition Award. He is also the recipient of the 2006 Lemuel J. Bowie Young Investigator Award for the Chicago Section of the AACC. Dr. Wang has authored over 140 journal articles, book chapters, and abstracts. He also serves on several editorial boards of peer reviewed journals.
After viewing this presentation, participants will be able to:
- Outline the challenges of antibody-based subtyping
- Recognize the higher analytical sensitivity and specificity of an LC/MS approach to protein subtyping
- Understand how LC/MS can be used to identify and subtype amyloid proteins
Clinical researchers who are interested in: aberrations in protein expression; translational proteomics; clinical research applications; study of disease mechanisms; antibody-based subtyping; and/or use of LC-MS tools in the clinical research lab.
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