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Toward Top-Down Determination of PEGylation Site Using MALDI In-Source Decay MS Analysis
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Toward Top-Down Determination of PEGylation Site Using MALDI In-Source Decay MS Analysis
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Journal Article
Toward Top-Down Determination of PEGylation Site Using MALDI In-Source Decay MS Analysis
2009
Overview
A novel matrix assisted laser desorption/ionization (MALDI)-based mass spectrometric approach has been evaluated to rapidly analyze a custom designed PEGylated peptide that is 31 residues long and conjugated with 20 kDa linear polyethylene glycol (PEG) at the side chain of Lys. MALDI-TOF MS provided sufficiently high resolution to allow observation of each of the oligomers of the heterogeneous PEGylated peptide (m/Δm of ca. 500), while a typical ESI-MS spectrum of this molecule was extremely complex and unresolved. Reflector in-source decay (reISD) analysis using MALDI-TOF MS was attempted to identify the PEGylation site at intact molecular level without any sample treatment. An reISD spectrum of the free peptide was observed with abundant c-, y-, and [z + 2]-fragment ion series, whereas, in the fragmented PEGylated peptide, the fragment ion series were truncated at the residue where PEG was attached. Therefore, a direct comparison of these top-down reISD spectra suggested the location of the PEGylation site. Results from this study demonstrate a clear analytical utility of the ISD technique to characterize structural aspects of heterogeneous biomolecules.
A MALDI-based in-source decay technique was utilized for rapid top-down MS analysis of a peptide conjugated with a 20 kDa polyethylene glycol.
Publisher
Elsevier Inc,Springer-Verlag,Elsevier,Springer Nature B.V
Subject
Aminoacids, peptides. Hormones. Neuropeptides
/ Analytical, structural and metabolic biochemistry
/ Biological and medical sciences
/ Chemistry and Materials Science
/ Decay
/ Fundamental and applied biological sciences. Psychology
/ Ions
/ Peptides
/ Polyethylene Glycols - chemistry
/ Proteins
/ Spectrometry, Mass, Electrospray Ionization - methods
/ Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization - methods
