De bepaling van het molecuulgewicht van Fab- en Fc-fragmenten van het eiwit IgG2a met Plasma Desorptie Massaspectrometrie en Electrospray Massaspectrometrie
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Open Access
Type
Report
Language
nl
Date
1990-07-31
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Title
De bepaling van het molecuulgewicht van Fab- en
Fc-fragmenten van het eiwit IgG2a met Plasma Desorptie Massaspectrometrie en
Electrospray Massaspectrometrie
Translated Title
The determination of the molecularweight of Fab-
and Fc-fragments of the albumen with Plasma Desorption Mass spectrometry and
Electrospray Mass spectrometry
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Abstract
Abstract niet beschikbaar
Recently, two mass spectrometric techniques have been introduced for the accurate determination of the molecular weight of proteins and peptides, namely Plasma Desorption-Mass Spectrometry (PD-MS) and Electro Spray- Mass Spectrometry (ES-MS). In this study the applicability of these techniques to laboratory samples has been tested. Measurements have been performed in the framework of the study on the stability of monoclonal antibodies (mabs) like IgG2a. Because the molecular weight of IgG2a exceeds the mass range of the PD-MS instrument, the test compound has been cleaved into Fab and Fc fragments with expected molecular weights after reduction of approximately 25,000 Dalton. The Fab and Fc fragments have been separated, reduced, solved in 0.01% sodium azide solution, and were offered for analysis as two separate samples. With the ES-MS technique in both samples two compounds have been detected with molecular weights in the range of 24,000 to 26,000 Dalton. The molecular weights were determined with an accuracy better than 0.02%. The sample amount used was 100-200 pmol. With the PD-MS technique no interpretable spectra have been obtained.
Recently, two mass spectrometric techniques have been introduced for the accurate determination of the molecular weight of proteins and peptides, namely Plasma Desorption-Mass Spectrometry (PD-MS) and Electro Spray- Mass Spectrometry (ES-MS). In this study the applicability of these techniques to laboratory samples has been tested. Measurements have been performed in the framework of the study on the stability of monoclonal antibodies (mabs) like IgG2a. Because the molecular weight of IgG2a exceeds the mass range of the PD-MS instrument, the test compound has been cleaved into Fab and Fc fragments with expected molecular weights after reduction of approximately 25,000 Dalton. The Fab and Fc fragments have been separated, reduced, solved in 0.01% sodium azide solution, and were offered for analysis as two separate samples. With the ES-MS technique in both samples two compounds have been detected with molecular weights in the range of 24,000 to 26,000 Dalton. The molecular weights were determined with an accuracy better than 0.02%. The sample amount used was 100-200 pmol. With the PD-MS technique no interpretable spectra have been obtained.
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