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In‐depth analysis of low abundant proteins in bovine colostrum using different fractionation techniques
Author(s) -
Nissen Asger,
Bendixen Emøke,
Ingvartsen Klaus Lønne,
Røntved Christine Maria
Publication year - 2012
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.201200231
Subject(s) - colostrum , fractionation , proteome , chromatography , chemistry , proteomics , biology , biochemistry , antibody , immunology , gene
Bovine colostrum is well known for its large content of bioactive components and its importance for neonatal survival. Unfortunately, the colostrum proteome is complicated by a wide dynamic range, because of a few dominating proteins that hamper sensitivity and proteome coverage achieved on low abundant proteins. Moreover, the composition of colostrum is complex and the proteins are located within different physical fractions that make up the colostrum. To gain a more exhaustive picture of the bovine colostrum proteome and gather information on protein location, we performed an extensive pre‐analysis fractionation of colostrum prior to 2 D ‐ LC ‐ MS / MS analysis. Physical and chemical properties of the proteins and colostrum were used alone or in combination for the separation of proteins. ELISA was used to quantify and verify the presence of proteins in colostrum. In total, 403 proteins were identified in the nonfractionated colostrum ( NF ) and seven fractions (F1‐F7) using six different fractionation techniques. Fractionation contributed with 69 additional proteins in the fluid phase compared with NF. Different fractionation techniques each resulted in detection of unique subsets of proteins. Whey production by high‐speed centrifugation contributed most to detection of low abundant proteins. Hence, prefractionation of colostrum prior to 2 D ‐ LC ‐ MS / MS analysis expanded our knowledge on the presence and location of low abundant proteins in bovine colostrum.

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