New Proteomic Subfractionation Surfaces - Innovative Technology For The Improved Resolution of Serum Proteins
Swapan Roy. Ph.D., John Krupey. Ph.D., Matthew Kuruc, Devjit Roy ProFACT Protomics, Inc.
Commercialization Center for Innovative Technologies
675 Route One
North Brunswick NJ 08902
Tel: 732-246-1190
Fax: 732-246-3118
Abstract
ProFACT™ is a new sorting, or subfractionation methodology designed for comparative
proteome analysis. Electrophoretic profiles of serum subfractions demonstrate improved resolution and
quantification. Carryover from the three highest abundance serum proteins, albumin, IgG and Transferrin
is minimal. The process starts with a separation platform utilizing a new combination of surface microenvironments
substituted with low molecular weight substrates that feature drug-binding motifs. With the ProFACT™
surface library, undenatured, bioactive proteins can be subfractionated into differential pools.
Separations are universal as they do not require pre-qualified binding knowledge, a key limitation of
affinity-type techniques. The surfaces utilized are disposable and versatile to meet sample size and scale
requirements. A simple bind, wash and elute protocol is completed in 30 to 60 minutes.
As elutions are mild and consistent, a direct handoff can be made to subsequent interrogation. The interrogation
strategy is adaptable to meet investigative inquiry using conventional reporter/probe bioassays, high performance
resolution or optical techniques. Subsequent patterns may have diagnostic potential as biomarkers, or be useful
for research-driven needs as a means to reduce the initial complexity of the crude sample down to a manageable number
of subfractions that show differences, the “hits”. These hits can then be further analyzed by HPLC, Capillary
Electrophoresis, 1 and 2D Electrophoresis, and Mass Spectrometry to more definitively identify and
characterize discoveries.
As structural modifications of proteins can alter their binding affinities to the surface library, differences
in structural features are revealed upon quantitative interpretation of ProFACT™ subfraction profiles.
Future investigations will focus on comparing normal and disease state sera.