Browse views: by Year, by Function, by GLF, by Subfunction, by Conference, by Journal

The use of generic surrogate peptides for the quantitative analysis of immunoglobulin G1 in pre-clinical species with high-resolution mass spectrometry

Lanshoeft, Christian and Wolf, Thierry and Heudi, Olivier and Cianferani, Sarah and Barteau, Samuel and Walles, Markus and Picard, Franck and Kretz, Olivier (2016) The use of generic surrogate peptides for the quantitative analysis of immunoglobulin G1 in pre-clinical species with high-resolution mass spectrometry. Analytical and bioanalytical Chemistry.

Abstract

In the present study, the application of a liquid chromatography high-resolution mass spectrometry (LC-HRMS) analytical assay for the quantitative analysis of a recombinant humanized immunoglobulin G1 (IgG1) in rat serum is reported using three generic peptides GPSVFPLAPSSK (GPS), TTPPVLDSDGSFFLYSK (TTP) and VVSVLTVLHQDWLNGK (VVS). Moreover, the deamidation site of a fourth peptide FNWYVDGVEVHNAK (FNW) was identified and further excluded from the assay evaluation due to the inaccuracy of the quantitative results. The rat serum samples were spiked with a fully labeled IgG1 as internal standard (ISTD). The digestion with trypsin was performed onto the pellet prior to peptide analysis by LC-HRMS using a quadrupole time of flight (QTOF) mass analyzer operating in selected reaction monitoring (SRM) mode with enhanced duty cycles (EDC). The assay linearity for the three investigated peptides was established for a humanized IgG1 (IgG1A) from 1.00 to 1000 µg mL-1 with a mean coefficient of determination (R2) higher than 0.9868. The inter-day accuracy and precision obtained in rat serum over three days was ≤11.4 % and ≤10.5 %, respectively. Short-term stability on the auto-sampler at 6 °C for 30 h, at RT for 48 h and a 100-fold dilution factor were demonstrated. In addition, QC samples prepared in cynomolgus monkey serum and measured with the present method met the acceptance criteria of ±20.0 % and ≤20.0 % for all three peptides regarding accuracy and precision, respectively. The LC-HRMS method was applied to the analysis of samples from five individual cynomolgus monkeys dosed with a second IgG1 (IgG1B) and consistent data were obtained compared to the LC-MS/MS method (conventional triple quadrupole (QqQ) mass analyzer operating in SRM). The present data demonstrate that LC-HRMS can be used for the quantitative analysis of IgG1 in both species and that quantification is not only limited to classical QqQ instruments.

Item Type: Article
Keywords: high-resolution mass spectrometry; generic peptide; immunoglobulin G; pellet digestion, pre-clinical species, deamidation
Date Deposited: 09 May 2016 23:45
Last Modified: 09 May 2016 23:45
URI: https://oak.novartis.com/id/eprint/27276

Search

Email Alerts

Register with OAK to receive email alerts for saved searches.