Hexadecanedionic Acid−Sepharose 4B:  A New Tool for Preparation of Albumin-Depleted Plasma

Abstract

Serum and plasma are the major sources of human material for clinical molecular diagnostics and drug discovery. However, due to the high abundance of some proteins, of which serum albumin (SA) is most prominent, lower-abundance proteins often remain undetectable in proteomic analysis of these body fluids. We have used hexadecanedionic acid (HDA) immobilized to Sepharose 4B to develop an affinity resin that is effective in the removal of SA from plasma. Two-dimensional gel analysis of the SA-depleted samples shows a significant enhancement of the low-abundance proteins and highly specific capture of serum albumin. The HDA resin shows better performance in terms of specificity than dye-based resins.

Depletion of High-Abundance Serum Proteins from Human Uterine Lavages Enhances Detection of Lower-Abundance Proteins

“…These include protein precipitation by trichloroacetic acid (TCA) , or a combination of TCA/acetone or TCA/ethanol. Several affinity matrices for abundant serum protein depletion have been developed including cibachron blue (a chlorotriazine dye) and hexadecanedionic acid immobilized to Sepharose resin, both of which have a high affinity for albumin and can effectively remove albumin from plasma, − and Protein A or G , which depletes immunoglobulin (Ig). However, such affinity materials, particularly those that are dye-based, often bind many other proteins nonspecifically.…”