Journal
PROTEOMICS
Volume 11, Issue 18, Pages 3633-3641Publisher
WILEY-BLACKWELL
DOI: 10.1002/pmic.201100023
Keywords
Immunoassay; MS; Plasma; Quantification; Technology; Transthyretin; Trypsin
Funding
- National Center for Research Resources (NCRR), National Institutes of Health (NIH) [1 R43 RR025701]
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Transthyretin (TTR, or prealbumin) is a tetrameric protein found in plasma and cerebrospinal fluid. Its major role is to transport thyroid hormones (thyroxin-T4) and retinol (through association with retinol-binding protein). TTR has been studied extensively due to the great number of point mutations that result in sequence heterogeneity. Many of these variants are associated with pathological conditions that result in extracellular deposition of amyloid fibers in tissues. In this work, we have developed a rapid mass spectrometric immunoassay for determination and quantification of TTR and its variants from human serum and plasma samples. The assay was fully characterized in terms of its precision, linearity and recovery characteristics. The new assay was also compared with a conventional TTR ELISA. Furthermore, we have applied the optimized method to analyze TTR and its modifications in 44 human plasma samples, and in the process optimized a method for TTR proteolytic digestion and identification of point mutations.
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