Polystyrene beads as an alternative support material for epitope identification of a prion-antibody interaction using proteolytic excision mass spectrometry

Pimenova, Tatiana; Meier, Lukas; Roschitzki, Bernd; Paraschiv, Gabriela-Ioana; Przybylski, Michael; Zenobi, Renato

Polystyrene beads as an alternative support material for epitope identification of a prion-antibody interaction using proteolytic excision mass spectrometry

Dateien

Polystyrene_pimenova.pdfGröße: 629.4 KBDownloads: 505

Datum

2009

Autor:innen

Pimenova, Tatiana

Meier, Lukas

Roschitzki, Bernd

Paraschiv, Gabriela-Ioana

Przybylski, Michael

Zenobi, Renato

Open Access-Veröffentlichung

Open Access Green

Sammlungen

Chemie

Publikationstyp

Zeitschriftenartikel

Publikationsstatus

Published

Erschienen in

Analytical and Bioanalytical Chemistry. 2009, 395(5), pp. 1395-1401. ISSN 1618-2642. eISSN 1618-2650. Available under: doi: 10.1007/s00216-009-3119-8

Zusammenfassung

The binding epitope structure of a protein specifically recognized by an antibody provides key information to prevent and treat diseases with therapeutic antibodies and to develop antibody-based diagnostics. Epitope structures of antigens can be effectively identified by the proteolytic epitope excision mass spectrometry (MS) method, which involves (1) immobilization of monoclonal or polyclonal antibodies, e.g., on N-hydroxysuccinimide-activated sepharose, (2) affinity binding of the antigen followed by limited proteolytic digestion of the
immobilized immune complex, and (3) elution and mass spectrometric analysis of the remaining affinity-bound peptide(s). In the epitope analysis of recombinant cellular bovine prion protein (bPrPC) to a monoclonal antibody (mAb3E7), we found that epitope excision experiments resulted in extensive nonspecific binding of bPrP to a standard sepharose matrix employed. Here, we show that the use of amino-modified polystyrene beads with aldehyde functionality is an efficient alternative support for antibody immobilization, suitable for epitope excision MS, with complete suppression of nonspecific bPrP binding.

Fachgebiet (DDC)

540 Chemie

Schlagwörter

Epitope excision, Polystyrene beads, Prion protein, Antibody antigen interaction, Mass spectrometry

Zitieren

ISO 690

PIMENOVA, Tatiana, Lukas MEIER, Bernd ROSCHITZKI, Gabriela-Ioana PARASCHIV, Michael PRZYBYLSKI, Renato ZENOBI, 2009. Polystyrene beads as an alternative support material for epitope identification of a prion-antibody interaction using proteolytic excision mass spectrometry. In: Analytical and Bioanalytical Chemistry. 2009, 395(5), pp. 1395-1401. ISSN 1618-2642. eISSN 1618-2650. Available under: doi: 10.1007/s00216-009-3119-8

BibTex

@article{Pimenova2009Polys-10002,
  year={2009},
  doi={10.1007/s00216-009-3119-8},
  title={Polystyrene beads as an alternative support material for epitope identification of a prion-antibody interaction using proteolytic excision mass spectrometry},
  number={5},
  volume={395},
  issn={1618-2642},
  journal={Analytical and Bioanalytical Chemistry},
  pages={1395--1401},
  author={Pimenova, Tatiana and Meier, Lukas and Roschitzki, Bernd and Paraschiv, Gabriela-Ioana and Przybylski, Michael and Zenobi, Renato}
}

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Universitätsbibliographie

Ja