Fig. 2.

Characterization of target peptide standards. a Standard curves for each target analyte peptide over a 9 point dilution showing linearity from 275 fmols to 1 fmol total GP₁. An aliquot of the previously quantified eVLP lot ‘A’ (200 fmols/µL SEE at 120 µL dilution) was resuspended 83 µL 40 % acetonitrile, 0.1 % Formic (137.5 fmols/µL) and serially diluted. A 2 µL injection utilizing the described instrument method was run in triplicate for each dilution. R² values for all four peptides are well within the margin of significance for linearity. Also shown in tabular form are the %CV values for each triplicate XIC measurement for each peptide at each dilution. These data indicate linearity down to 1 fmol with the largest CV% (SVGR—17.3 %) in dilution number ‘8’ of the serially diluted series. b AQUA-SVG peptide signal response for non-deamidated (circle) and deamidated (triangle) peptide. AQUA-SVG peptide was deamidated by incubating 40 pmols at 50 °C/pH 8.0 for 2.5 days while a matching 40 pmol aliquot was stored at −20 °C. A 5-point, twofold serial dilution was performed resulting in a 250–15.6 fmol/µL concentration range for each sample. LC-HRMS was run in triplicate on each dilution and the average counts plotted