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    Development of sensitive mass spectrometric techniques in protein and peptide identification

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    Authors
    de Jong APJM
    Meiring HD
    Barosso B
    ten Hove GJ
    van der Heeft E
    Type
    Report
    Language
    en
    
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    Title
    Development of sensitive mass spectrometric techniques in protein and peptide identification
    Translated Title
    Ontwikkeling van gevoelige massa spectrometrische technieken voor de identificatie van eiwitten en peptiden
    Publiekssamenvatting
    Dit rapport beschrijft de ontwikkeling van micro en nano-scale vloeistofchromatografie en on-line preconcentrering capillaire electrophorese gecombineerd met tandem massaspectrometrie bij gevoelige structuuranalyse analyse van eiwitten en peptiden. Massaspectrometrie is een sterk opkomende techniek voor het verkrijgen van sekwentie-informatie van deze biologisch relevante producten. De verkregen sekwentie-informatie van afgeleide peptiden (proteosomale peptiden en proteolyse producten) worden gebruikt voor het identificeren van het bijbehorende broneiwit met behulp van eiwit en DNA databanken. De gevoeligheid van de huidige MS technologieen ligt rond een picomol (10-12) hoeveelheid eiwit. Recente verfijningen, in het bijzonder de ontwikkeling van nano-schaal technologien brengen toepassingen op het femto (10-15) tot attomole (10-18) niveau binnen het bereik van de techniek. Hier rapporteren wij over de resultaten van het R&D programma in het jaar 1998 op het gebied van de karakterisering van eiwitten en peptiden met behulp van massaspectrometrie.<br>
    This report describes the development of micro- and nano-scale liquid chromatographic and capillary zone electrophoretic separation techniques, combined with tandem mass spectrometry, for the sensitive structure analysis of proteins and peptides. Mass spectrometry is emerging as the method of choice for the characterisation and identification of unknown proteins and peptides. Sequence information obtained from derived peptides, such as naturally processed antigenic peptides and proteolytic fragments is used to identify the underlying source protein using an online database search for protein and DNA databases. Typical sensitivities of current MS methods come to around the sub-pmol (10-12) level of protein on a routine basis. Ongoing refinements in the technology, particularly the development of nano-scale technologies, makes successful applications possible down to the femto (10-15) and attomole (10-18) levels. Here we report on the results this laboratory will have developments in this field over the past year.<br>
    Publisher
    Rijksinstituut voor Volksgezondheid en Milieu RIVM
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