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Run on QSTAR Elite, November 2008. lactoferrin. invertase. All samples PNGase treated. Depl Plasma + 500 pmol GPS, Depl Plasma + 100 pmol GPS, Depl Plasma + 50 pmol GPS, Depl Plasma + 10 pmol GPS, Depl Plasma + 1 pmol GPS, Depl Plasma + 500 fmol GPS,. Protein hit #1. Protein hit #7.
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Run on QSTAR Elite, November 2008 lactoferrin invertase All samples PNGase treated • Depl Plasma + 500 pmol GPS, • Depl Plasma + 100 pmol GPS, • Depl Plasma + 50 pmol GPS, • Depl Plasma + 10 pmol GPS, • Depl Plasma + 1 pmol GPS, • Depl Plasma + 500 fmol GPS, Protein hit #1 Protein hit #7 Protein hit #6 Protein hit #37 Protein hit #6 Protein hit #39 Protein hit #54 Protein hit #62 Not observed Not observed Not observed Not observed At 10 pmol we first see lactoferrin and invertase as protein hit. The deamidated target peptides then show up at 50, 100, 500 pmol (see next slide)
Lacto(trans)ferrin – positive control (spiked GPS in plasma) Run on QSTAR Elite, November 2008 100 pmol 500 pmol 50 pmol score 96 score 45 score 30 R.TAGWNIPMGLLFNQTGSCK.F score 99 GLLFNQTGSCK.F score 99 Tryptic cleavage occurs score 98 R.TAGWNVPIGTLRPFLNWTGPPEPIEAAVAR.F LNWTGPPEPIEAAVAR.F score 95 PFLNWTGPPEPIEAAVAR.F score 75 score 99 score 97 score 99 score 99 R.TAGWNVPIGTLR score 99 R.TAGWNVPIGTLRPF LNWTGPPEPIEAAVAR.F N deamidated Asn Search scores from Protein Pilot
Run on QSTAR Elite, November 2008 • 1/10th of 100 ul plasma + 500 pmol GPS, PNGaseF-treated • (081125_0008_PD#1_lectin.wiff) • 1/10th of 100 ul plasma + 100 pmol GPS, PNGaseF-treated • (081125_0007_PD#2_lectin.wiff) • 1/10th of 100 ul plasma + 50 pmol GPS, PNGaseF-treated • 081125_0006_PD#3_lectin.wiff • 1/10th of 100 ul plasma + 10 pmol GPS, PNGaseF-treated • 081125_0005_PD#4_lectin.wiff • 1/10th of 100 ul plasma + 1 pmol GPS, PNGaseF-treated • 081125_0004_PD#5_lectin.wiff • 1/10th of 100 ul plasma + 500 fmol GPS, PNGaseF-treated • 081125_0003_PD#6_lectin.wiff.scan