Hi3 peptide quantitative analysis of abundant plasma proteins.

Median (ng/mL): median of protein concentrations determined from number of samples shown in column n. IRV: inter-assay variation, the coefficients of variation obtained from 7 aliquots of a pooled sample separately digested and measured over the course of 3 months of normal operation of the instrument. IAV: intra-assay variation, coefficient of variation obtained from 6 aliquots of a pooled sample separately digested and subsequently measured during 1 day. AV: analytical variability, determined from 10 replicate injections of a single digested sample throughout 9 days of measurements. LIN: linearity of measurements, the Pearson’s correlation coefficient shows linearity between total protein load on the analytical column and nanogram protein quantified by HI3 peptide quantitation using the ADH1 digest standard; only determined when at least 4 points were available for a protein. See also S1 Fig. ST: Pearson’s correlation of protein quantitation using two different digest standards (ADH1 and ENO1 from yeast). LOQ: limit of quantitation (ng/mL) estimated by dilution of a plasma sample in a constant background of a digest standard. n: number of samples (out of 31) in which the protein was quantified, FN: protein number on the x-axis of Fig 1c. hc: heavy chain.
(50:1 substrate:enzyme, 20 hours digestion, 0.21 ug on column injection).