Immunoaffinity fractionation of plasma proteins by chicken IgY antibodies.

Separation of complex mixtures having a wide dynamic range of protein concentration, such as plasma or serum, presents a significant challenge for proteomic analysis. Immunoaffinity fractionation is one of the most effective methods used during sample preparation to improve the ability to detect low-abundant proteins (LAP), enhancing biomarker discovery. Avian IgY (Immunoglobulin Yolk) antibodies have unique and advantageous features, which include strong avidity, high specificity, low nonspecific binding, and accumulative production. Polyclonal IgY antibodies covalently coupled to microbeads are particularly effective in specifically removing high-abundant proteins (HAP) from plasma, serum, CSF, urine, and other body fluid or cellular sources. IgY-12 is a composition of IgY microbeads designed for one-step removal of the 12 most abundant proteins in human serum or plasma: albumin, IgG, transferrin, fibrinogen, alpha1-antitrypsin, IgA, IgM, alpha2-macroglobulin, haptoglobin, apolipoproteins A-I and A-II, and orosomucoid (alpha1-acid glycoprotein). Removal of the 12 HAPs enables improved resolution and dynamic range for one-dimensional gel electrophoresis (1DGE), two-dimensional gel electrophoresis (2DGE), and liquid chromatography/mass spectrometry (LC/MS).