Peptide storage: are you getting the best return on your investment? Defining optimal storage conditions for proteomics samples.
To comply with current proteomics guidelines, it is often necessary to analyze the same peptide samples several times. Between analyses, the sample must be stored in such a way as to conserve its intrinsic properties, without losing either peptides or signal intensity. This article describes two studies designed to define the optimal storage conditions for peptide samples between analyses. With the use of a label-free strategy, peptide conservation was compared over a 28-day period in three different recipients: standard plastic tubes, glass tubes, and low-adsorption plastic tubes. The results of this study showed that standard plastic tubes are unsuitable for peptide storage over the period studied. Glass tubes were found to perform better than standard plastic, but optimal peptide recovery was achieved using low-adsorption plastic tubes. The peptides showing poor recovery following storage were mainly hydrophobic in nature. The differences in peptide recovery between glass and low-adsorption plastic tubes were further studied using isotopically labeled proteins. This study allowed accurate comparison of peptide recovery between the two tube types within the same LC-MS run. The results of the label-free study were confirmed. Further, it was possible to demonstrate that peptide recovery in low-adsorption plastic tubes was optimal whatever the peptide concentration stored.
Kraut
Alexandra
A
CEA, DSV, iRTSV, Laboratoire d'Etude de la Dynamique des Protéomes, INSERM, U880, Université Joseph Fourier, Grenoble, F-38054, France.
Marcellin
Marlène
M
Adrait
Annie
A
Kuhn
Lauriane
L
Louwagie
Mathilde
M
Kieffer-Jaquinod
Sylvie
S
Lebert
Dorothée
D
Masselon
Christophe D
CD
Dupuis
Alain
A
Bruley
Christophe
C
Jaquinod
Michel
M
Garin
Jérôme
J
Gallagher-Gambarelli
Maighread
M
D000327:Adsorption; D000818:Animals; D002417:Cattle; D004926:Escherichia coli / Q000378:metabolism; D013058:Mass Spectrometry / Q000379:methods; D010455:Peptides / Q000737:chemistry*; D011506:Proteins / Q000737:chemistry; D040901:Proteomics / Q000295:instrumentation* / Q000379:methods*; D011817:Rabbits; D013048:Specimen Handling; D013194:Staining and Labeling; D013997:Time Factors
D010455:Peptides; D011506:Proteins