new method for prefractionation of plasma for proteomic analysis
TRANSCRIPT
This article was originally published in Electrophoresis 2010, 31, 3580–3585, DOI 10.1002/elps.201000298
New method for prefractionation of plasma for
proteomic analysis
Anna Fitzgerald and Bradley J. Walsh
Keywords:
2-DE / IEF / Plasma / Prefractionation
The depth of proteome analysis is severely limited in
complex samples with a wide dynamic range of protein
abundance such as plasma. Removal of high-abundance
proteins should reveal the signal of lower abundance plasma
proteins. However, smaller proteins may be part of larger
protein complexes and hence the removal of proteins
involved in complexes with high-abundance proteins such
as albumin may inhibit the search for disease biomarkers.
Prefractionation of a sample divides it into fractions of
reduced complexity, allowing improved detection of lower
abundance proteins. Using a prefractionation device, which
employs GradiflowTM technology, we were able to separate
small volume plasma samples into multiple fractions based
on the molecular weight and/or charge. The resulting
samples of reduced complexity were directly compatible
with 2-DE. The use of this prefractionation machine may
therefore be useful in the hunt for disease biomarkers.
Time course experiment showing native separation of plasmaafter fractionation based on MW in the MF-10 Gradiflow.
Proteomics Clin. Appl. 2011, 5, 205–209 209