The minotaur proteome: avoiding cross-species identifications deriving from bovine serum in cell culture models.
Cell culture is a fundamental tool in proteomics where mammalian cells are cultured in vitro using a growth medium often supplemented with 5-15% FBS. Contamination by bovine proteins is difficult to avoid because of adherence to the plastic vessel and the cultured cells. We have generated peptides from bovine serum using four sample preparation methods and analyzed the peptides by high mass accuracy LC-MS/MS. Distinguishing between bovine and human peptides is difficult because of a considerable overlap of identical tryptic peptide sequences. Pitfalls in interpretation, different database search strategies to minimize erroneous identifications and an augmented contaminant database are presented.
Bunkenborg
Jakob
J
Department of Biochemistry and Molecular Biology, University of Southern Denmark, Odense M, Denmark.
García
Guadalupe Espadas
GE
Paz
Marcia Ivonne Peña
MI
Andersen
Jens S
JS
Molina
Henrik
H
D000818:Animals; D016477:Artifacts; D002417:Cattle; D018929:Cell Culture Techniques / Q000379:methods*; D002478:Cells, Cultured; D003470:Culture Media / Q000737:chemistry*; D004340:Drug Contamination; D006801:Humans; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism; D020543:Proteome / Q000737:chemistry* / Q000378:metabolism; D040901:Proteomics / Q000592:standards*; D044967:Serum / Q000737:chemistry*; D013045:Species Specificity; D014357:Trypsin / Q000378:metabolism
D003470:Culture Media; D010446:Peptide Fragments; D020543:Proteome; D014357:Trypsin