Practical immunoaffinity-enrichment LC-MS for measuring protein kinetics of low-abundance proteins.

ABSTRACT:

Background

For a more complete understanding of pharmacodynamic, metabolic, and pathophysiologic effects, protein kinetics, such as production rate and fractional catabolic rate, can offer substantially more information than protein concentration alone. Kinetic experiments with stable isotope tracers typically require laborious sample preparation and are most often used for studying abundant proteins. Here we describe a practical methodology for measuring isotope enrichment into low-abundance proteins that uses an automated procedure and immunoaffinity enrichment (IA) with LC-MS. Low-abundance plasma proteins cholesteryl ester transfer protein (CETP) and proprotein convertase subtilisin/kexin type 9 (PCSK9) were studied as examples.

Methods

Human participants (n = 39) were i

SUBMITTER: Lassman ME 

PROVIDER: S-EPMC4955776 | biostudies-literature | 2014 Sep

REPOSITORIES: biostudies-literature