Protein Equalizer Technology : the quest for a "democratic proteome".
No proteome can be considered "democratic", but rather "oligarchic", since a few proteins dominate the landscape and often obliterate the signal of the rare ones. This is the reason why most scientists lament that, in proteome analysis, the same set of abundant proteins is seen again and again. A host of pre-fractionation techniques have been described, but all of them, one way or another, are besieged by problems, in that they are based on a "depletion principle", i.e. getting rid of the unwanted species. Yet "democracy" calls not for killing the enemy, but for giving "equal rights" to all people. One way to achieve that would be the use of "Protein Equalizer Technology" for reducing protein concentration differences. This comprises a diverse library of combinatorial ligands coupled to spherical porous beads. When these beads come into contact with complex proteomes (e.g. human urine and serum, egg white, and any cell lysate, for that matter) of widely differing protein composition and relative abundances, they are able to "equalize" the protein population, by sharply reducing the concentration of the most abundant components, while simultaneously enhancing the concentration of the most dilute species. It is felt that this novel method could offer a strong step forward in bringing the "unseen proteome" (due to either low abundance and/or presence of interference) within the detection capabilities of current proteomics detection methods. Examples are given of equalization of human urine and serum samples, resulting in the discovery of a host of proteins never reported before. Additionally, these beads can be used to remove host cell proteins from purified recombinant proteins or protein purified from natural sources that are intended for human consumption. These proteins typically reach purities of the order of 98%: higher purities often become prohibitively expensive. Yet, if incubated with "equalizer beads", these last impurities can be effectively removed at a small cost and with minute losses of the main, valuable product.
Righetti
Pier Giorgio
PG
Department of Chemistry, Materials and Chemical Engineering Giulio Natta, Politecnico di Milano, Milano, Italy. piergiorgio.righetti@polimi.it
Boschetti
Egisto
E
Lomas
Lee
L
Citterio
Attilio
A
D015415:Biomarkers / Q000737:chemistry; D001798:Blood Proteins / Q000737:chemistry / Q000302:isolation & purification; D004274:DNA, Recombinant / Q000737:chemistry; D006801:Humans; D008024:Ligands; D008863:Microspheres; D019151:Peptide Library; D040901:Proteomics / Q000379:methods*; D014556:Urine / Q000737:chemistry
D015415:Biomarkers; D001798:Blood Proteins; D004274:DNA, Recombinant; D008024:Ligands; D019151:Peptide Library