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

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Author(s) Pimenova, Tatiana, Meier, Lukas, Roschitzki, Bernd, Paraschiv, Gabriela, Przybylski, Michael, Zenobi, Renato
Publication Type Journal Items, Publication Status: Published
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Title Polystyrene beads as an alternative support material for epitope identification of a prion-antibody interaction using proteolytic excision–mass spectrometry
Author(s) Pimenova, Tatiana
Meier, Lukas
Roschitzki, Bernd
Paraschiv, Gabriela
Przybylski, Michael
Zenobi, Renato
Journal or Series Title Analytical and bioanalytical chemistry
Volume Number 395
Issue Number 5
Start Page 1395
End Page 1401
ISSN 1618-2642
1618-2650
Publisher Springer
Publication Place Berlin
Publication Date 2009
Keyword(s) Epitope excision
Polystyrene beads
Prion protein
Antibody-antigen interaction
Mass spectrometry
Abstract 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 (bPrP(C)) 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.
DOI 10.1007/s00216-009-3119-8
Additional Notes Received 7 June 2009, Revised 24 August 2009, Accepted 29 August 2009, Published online 30 September 2009
Document Type Article
Publication Status Published
Language English
Assigned Organisational Unit(s) 02207
03430
Organisational Unit(s)
NEBIS System Number 004239219
Source Database ID PP-49612
WOS-000270897000021
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@article{Pmnv2009,
  author = "Pimenova, Tatiana and Meier, Lukas and Roschitzki, Bernd and Paraschiv, Gabriela and Przybylski, Michael and Zenobi, Renato",
  title = "{P}olystyrene beads as an alternative support material for epitope identification of a prion-antibody interaction using proteolytic excision{\textendash}mass spectrometry",
  journal = "Analytical and bioanalytical chemistry",
  year = 2009,
  volume = "395",
  number = "5",
  pages = "1395--1401",
}


E-Citations record created: Tue, 04 May 2010, 12:04:45 CET