A key challenge in working with serum/plasma samples is the difficulty in observing low abundance proteins. It is estimated that 12 proteins (albumin, total IgG, α-1-antitrypsin, IgA, IgM, transferrin, haptoglobin, α-1-acid glycoprotein (orosomucoid), α-2-macroglobulin, HDL (apolipoproteins A-I & A-II) and fibrinogen) compose 96% of the protein mass in plasma (1). Immunoaffinity partitioning of these highly-abundant proteins (HAP) has proven to be an effective approach for enabling the detection of low abundance proteins (LAP). Hence, in order to delve deeper into the serum/plasma proteome, it is necessary to perform immunoaffinity depletion as a pre-fractionation step.
- 250μl of serum/plasma is required for both the IgY14 and Supermix column immunoaffinity partitioning (GenWay Biotech. Inc.)
- 50μl is required for the IgY14 immunoaffinity portioning only
- Anderson, N.L., Anderson, N.G. (2002) .The human plasma proteome: history, character, and diagnostic prospects. Mol. Cell Proteomics 1:845–867.
- Huang, L., Harvie, G., Feitelson, J., Gramatikoff, K., Herold, D., Allen, D., Amunngama, R., Hagler, R., Pisano, M., Zhang, W., and Fang, X. (2005) Immunoaffinity separation of plasma protein by IgY microbeads: Meeting the
needs of proteomic sample preparation and analysis. Proteomics, 5:3314-3328.