Characterization of antibody–antigen interactions

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Author(s) Bich, Claudia, Scott, Mike, Panagiotidis, Andreas, Wenzel, Ryan J., Nazabal, Alexis, Zenobi, Renato
Publication Type Journal Items, Publication Status: Published
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Title Characterization of antibody–antigen interactions
Subtitle Comparison between surface plasmon resonance measurements and high-mass matrix-assisted laser desorption/ionization mass spectrometry
Author(s) Bich, Claudia
Scott, Mike
Panagiotidis, Andreas
Wenzel, Ryan J.
Nazabal, Alexis
Zenobi, Renato
Journal or Series Title Analytical biochemistry
Volume Number 375
Issue Number 1
Start Page 35
End Page 45
ISSN 0003-2697
Publisher Elsevier
Publication Place San Diego
Publication Date 2008
Keyword(s) Matrix-assisted laser desorption/ionization (MALDI)
Surface plasmon resonance (SPR)
Antibody–antigen interaction
Abstract The interaction between the bovine prion protein (bPrP) and a monoclonal antibody, 1E5, was studied with high-mass matrix-assisted laser desorption/ionization (MALDI) mass spectrometry (MS) and surface plasmon resonance (SPR). In the case of MS a cross-linking stabilization was used prior to the analysis, whereas for SPR the antibody was immobilized and bPrP was injected. We compared the determination of parameters such as the epitope, the kinetics and binding strength, and the capacity of the antigen to bind two different antibodies. The two methods are highly complementary. SPR measurements require a lower amount of sample but are more time-consuming due to all of the necessary side steps (e.g., immobilization, regeneration). High-mass MALDI MS needs a higher overall amount of sample and cannot give direct access to the kinetic constants, but the analysis is faster and easier compared with SPR.
DOI 10.1016/j.ab.2007.11.016
Additional Notes Received 3 October 2007, Available online 17 November 2007
Document Type Article
Publication Status Published
Language English
Assigned Organisational Unit(s) 03430
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NEBIS System Number 000032007
Source Database ID PP-40960
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  author = "Bich, Claudia and Scott, Mike and Panagiotidis, Andreas and Wenzel, Ryan J. and Nazabal, Alexis and Zenobi, Renato",
  title = "{C}haracterization of antibody{\textendash}antigen interactions: {C}omparison between surface plasmon resonance measurements and high-mass matrix-assisted laser desorption/ionization mass spectrometry",
  journal = "Analytical biochemistry",
  year = 2008,
  volume = "375",
  number = "1",
  pages = "35--45",

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