Differential recovery of peptides from sample tubes and the reproducibility of quantitative proteomic data.
Differential recovery of peptides due to nonspecific adsorption can seriously compromise reproducibility and quality of proteomic data for peptide analyses by liquid chromatography-mass spectrometry (LC-MS). This study demonstrates large variations in reproducibility and quantitation of LC-MS data for peptides derived from tryptic digests of BSA upon storage in different sample tubes. Notably, we show that highly improved consistency and lower errors in quantitation of BSA tryptic peptides in replicate measurements is achieved with low-retention tubes compared to regular eppendorf tubes. Furthermore, qualitative differences in peptides detected by LC-MS were observed in the two types of storage tubes. These results illustrate the necessity for careful evaluation of storage vessels and conditions to minimize variability in sample quality for LC-MS experiments.
Bark
Steven J
SJ
The Skaggs School of Pharmacy and Pharmaceutical Sciences, The University of California, San Diego, 9500 Gilman Drive, MC 0744, La Jolla, California 92093-0744, USA. sbark@ucsd.edu
Hook
Vivian
V
D000327:Adsorption; D000818:Animals; D002417:Cattle; D002853:Chromatography, Liquid / Q000379:methods; D030562:Databases, Protein; D013058:Mass Spectrometry / Q000379:methods; D010455:Peptides / Q000737:chemistry*; D011506:Proteins / Q000737:chemistry; D020543:Proteome; D040901:Proteomics / Q000379:methods*; D015203:Reproducibility of Results; D012709:Serum Albumin / Q000737:chemistry; D014357:Trypsin / Q000737:chemistry
D010455:Peptides; D011506:Proteins; D020543:Proteome; D012709:Serum Albumin; D014357:Trypsin