Clinical Applications of LC-MS: Development and Research Application of a Highly Sensitive LC-MS Method for Quantification of a Cholesterol Protein in Plasma

Recorded on May 15, 2014


Immunoassays and LC–MS-MS have emerged as the two main approaches for quantifying peptides and proteins in biological samples. ELISA kits are available for quantification, but inherently lack the discriminative power to resolve isoforms and post-translational modifications (PTMs).

To address this issue, and given the complexity and wide dynamic range of the plasma proteome, we have developed and applied a mass spectrometry immunoassay–selected reaction monitoring (MSIA-SRM) research method to quantify PCSK9. Proprotein convertase subtilisin/kexin type 9 (PCSK9) is a key player in the regulation of circulating low density lipoprotein cholesterol (LDL-C). Both the distinct forms observed in plasma and PTMs of this protein that have been described in cell-based studies are likely to affect its function and thereby LDL-C levels.

This webinar will discuss this method, from method development (optimal antibody determination using MSIA and SRM LC–MS-MS in terms of PTM determination, LOD/LOQ, and linearity over PCSK9’s clinical dynamic range) to applying to a test case with control plasma samples and samples with a known mutation to determine method specificity. In summary, a robust and sensitive MSIA-SRM method was developed for the absolute quantification of all PCSK9 domains and a PTM in plasma.

Moderator:

Steve Brown

Technical Editor
LCGC

Speaker:

Joëlle R Pérusse, PhD

Postdoctoral Fellow
Institut de Recherches Cliniques de Montréal


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