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MALDI linear TOF mass spectrometry of PEGylated (glyco)proteins
Author(s) -
Seyfried Birgit K.,
Siekmann Jürgen,
Belgacem Omar,
Wenzel Ryan J.,
Turecek Peter L.,
Allmaier Günter
Publication year - 2010
Publication title -
journal of mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.475
H-Index - 121
eISSN - 1096-9888
pISSN - 1076-5174
DOI - 10.1002/jms.1746
Subject(s) - chemistry , pegylation , mass spectrometry , matrix assisted laser desorption/ionization , human serum albumin , molecular mass , chromatography , recombinant dna , analytical chemistry (journal) , desorption , biochemistry , polyethylene glycol , adsorption , organic chemistry , enzyme , gene
PEGylation of proteins is a fast growing field in biotechnology and pharmaceutical sciences owing to its ability to prolong the serum half‐life time of recombinant proteins. Matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry (MALDI TOF MS) has been shown to be a powerful tool in the analysis of several PEGylated small proteins. Here we present data obtained with a standard secondary electron multiplier (SEM) and a high mass (HM) detector combined with a MALDI linear TOF MS system for the detection of PEGylated (glyco)proteins in the range of 60–600 kDa. Examples of MALDI TOF MS of small (interferon α2a), middle (human serum albumin (HSA)) and high molecular mass proteins (coagulation factor VIII and von Willebrand factor (vWF), both heavily glycosylated proteins) are presented. The particular challenge for the analysis was the heterogeneity of the (glyco)proteins in the high molecular weight range in combination with additional PEGylation, which even introduced more heterogeneity and was more challenging for interpretation. Nevertheless, the performance of MALDI linear TOF MS with both detector systems in terms molecular weight and heterogeneity determination depending on the m / z range was superior to the other methods. Although the SEM was able to obtain information about protein PEGylation in the mass range up to 100 kDa (e.g. PEGylated HSA), the HM system was crucial for detection of HM ions (e.g. PEGylated recombinant vWF), which was impossible with the standard SEM. Copyright © 2010 John Wiley & Sons, Ltd.