title: Affinity separation and enrichment methods in proteomic analysis.
abstract: Protein separation or enrichment is one of the rate-limiting steps in proteomic studies. Specific capture and removal of highly-abundant proteins (HAP) with large sample-handling capacities are in great demand for enabling detection and analysis of low-abundant proteins (LAP). How to grasp and enrich these specific proteins or LAP in complex protein mixtures is also an outstanding challenge for biomarker discovery and validation. In response to these needs, various approaches for removal of HAP or capture of LAP in biological fluids, particularly in plasma or serum, have been developed. Among them, immunoaffinity subtraction methods based upon polyclonal IgY or IgG antibodies have shown to possess unique advantages for proteomic analysis of plasma, serum and other biological samples. In addition, other affinity methods that use recombinant proteins, lectins, peptides, or chemical ligands have also been developed and applied to LAP capture or enrichment. This review discusses in detail the need to put technologies and methods in affinity subtraction or enrichment into a context of proteomic and systems biology as "Separomics" and provides a prospective of affinity-mediated proteomics. Specific products, along with their features, advantages, and disadvantages will also be discussed.
authors.affiliation: Acrotics Corporation, San Diego, CA 92130 USA. xfang@acrotics.com
authors.forename: Xiangming
authors.initials: X
authors.lastname: Fang
authors.forename: Wei-Wei
authors.initials: WW
authors.lastname: Zhang
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000737:chemistry; D001798:Blood Proteins / Q000737:chemistry; D002846:Chromatography, Affinity / Q000379:methods*; D006801:Humans; D007074:Immunoglobulin G / Q000737:chemistry; D007136:Immunoglobulins / Q000737:chemistry; D008024:Ligands; D051379:Mice; D040901:Proteomics / Q000379:methods*; D011994:Recombinant Proteins / Q000737:chemistry; D015203:Reproducibility of Results; D049490:Systems Biology
chemical_list: D015415:Biomarkers; D001798:Blood Proteins; C016512:IgY; D007074:Immunoglobulin G; D007136:Immunoglobulins; D008024:Ligands; D011994:Recombinant Proteins