Peptide mapping analysis of mouse/human chimeric anti-CD20 antibody Rituximab produced in glycoengineered Nicotiana benthamiana plants
Description
These are the mass spectrometry data, whose analysis is detailed in "Plant-related Quality Attributes Affecting FcγRIIIa Binding: Affinity Chromatography Analysis of Rituximab Glycovariants from Nicotiana benthamiana" (Frontiers in Plant Science). The MS data were obtained from the characterization of the mouse/human chimeric anti-CD20 antibody Rituximab, produced in glycoengineered Nicotiana benthamiana plants. The different samples were subjected to tryptic digestion, and the resulting peptide mixtures were first analyzed using an X500 QTOF instrument after reverse-phase (RP) separation, to assess sequence coverage and the relative abundance of oxidized variants. The same samples were also analyzed by HILIC separation coupled with detection on an LTQ instrument to identify glycopeptides and determine their relative abundances. Additionally, plant-derived mAbs were separated by FcγRIIIa receptor-based affinity chromatography. Different fractions were collected, digested with trypsin, and analyzed by RPLC-MS/MS peptide mapping using an Exactive Orbitrap instrument.
Files
Steps to reproduce
The spectra can be opened using the corresponding software. Peptide mapping data acquired on the X500 QTOF were processed using Explorer for SCIEX OS (SCIEX). Data acquired on the LTQ instrument were analyzed by manual evaluation of MS/MS spectra and manual extraction and integration of glycopeptide-related peaks. Peptide mapping data from affinity chromatography fractions were processed using MaxQuant and manually elaborated.
Institutions
Categories
Funding
Italian Ministry of Education, Universities and Research