A global benchmark study using affinity-based biosensors

Rebecca L. Rich, Giuseppe A. Papalia, Peter J. Flynn, Jamie Furneisen, John Quinn, Joshua S. Klein, Phini S. Katsamba, M. Brent Waddell, Michael Scott, Joshua Thompson, Judie Berlier, Schuyler Corry, Mireille Baltzinger, Gabrielle Zeder-Lutz, Andreas Schoenemann, Anca Clabbers, Sebastien Wieckowski, Mary M. Murphy, Phillip Page, Thomas E. RyanJay Duffner, Tanmoy Ganguly, John Corbin, Satyen Gautam, Gregor Anderluh, Andrej Bavdek, Dana Reichmann, Satya P. Yadav, Eric Hommema, Ewa Pol, Andrew Drake, Scott Klakamp, Trevor Chapman, Dawn Kernaghan, Ken Miller, Jason Schuman, Kevin Lindquist, Kara Herlihy, Michael B. Murphy, Richard Bohnsack, Bruce Andrien, Pietro Brandani, Danny Terwey, Rohn Millican, Ryan J. Darling, Liann Wang, Quincy Carter, Joe Dotzlaf, Jacinto Lopez-Sagaseta, Islay Campbell, Paola Torreri, Sylviane Hoos, Patrick England, Yang Liu, Yasmina Abdiche, Daniel Malashock, Alanna Pinkerton, Melanie Wong, Eileen Lafer, Cynthia Hinck, Kevin Thompson, Carmelo Di Primo, Alison Joyce, Jonathan Brooks, Federico Torta, Anne Birgitte Bagge Hagel, Janus Krarup, Jesper Pass, Monica Ferreira, Sergei Shikov, Malgorzata Mikolajczyk, Yuki Abe, Gaetano Barbato, Anthony M. Giannetti, Ganeshram Krishnamoorthy, Bianca Beusink, Daulet Satpaev, Tiffany Tsang, Eric Fang, James Partridge, Stephen Brohawn, James Horn, Otto Pritsch, Gonzalo Obal, Sanjay Nilapwar, Ben Busby, Gerardo Gutierrez-Sanchez, Ruchira Das Gupta, Sylvie Canepa, Krista Witte, Zaneta Nikolovska-Coleska, Yun Hee Cho, Roberta D'Agata, Kristian Schlick, Rosy Calvert, Eva M. Munoz, Maria Jose Hernaiz, Tsafir Bravman, Monica Dines, Min Hsiang Yang, Agnes Puskas, Erica Boni, Jiejin Li, Martin Wear, Asya Grinberg, Jason Baardsnes, Olan Dolezal, Melicia Gainey, Henrik Anderson, Jinlin Peng, Mark Lewis, Peter Spies, Quyhn Trinh, Sergei Bibikov, Jill Raymond, Mohammed Yousef, Vidya Chandrasekaran, Yuguo Feng, Anne Emerick, Suparna Mundodo, Rejane Guimaraes, Katy McGirr, Yue Ji Li, Heather Hughes, Hubert Mantz, Rostislav Skrabana, Mark Witmer, Joshua Ballard, Loic Martin, Petr Skladal, George Korza, Ite Laird-Offringa, Charlene S. Lee, Abdelkrim Khadir, Frank Podlaski, Phillippe Neuner, Julie Rothacker, Ashique Rafique, Nico Dankbar, Peter Kainz, Erk Gedig, Momchilo Vuyisich, Christina Boozer, Nguyen Ly, Mark Toews, Aykut Uren, Oleksandr Kalyuzhniy, Kenneth Lewis, Eugene Chomey, Brian J. Pak, David G. Myszka

Research output: Contribution to journalArticlepeer-review

68 Scopus citations

Abstract

To explore the variability in biosensor studies, 150 participants from 20 countries were given the same protein samples and asked to determine kinetic rate constants for the interaction. We chose a protein system that was amenable to analysis using different biosensor platforms as well as by users of different expertise levels. The two proteins (a 50-kDa Fab and a 60-kDa glutathione S-transferase [GST] antigen) form a relatively high-affinity complex, so participants needed to optimize several experimental parameters, including ligand immobilization and regeneration conditions as well as analyte concentrations and injection/dissociation times. Although most participants collected binding responses that could be fit to yield kinetic parameters, the quality of a few data sets could have been improved by optimizing the assay design. Once these outliers were removed, the average reported affinity across the remaining panel of participants was 620 pM with a standard deviation of 980 pM. These results demonstrate that when this biosensor assay was designed and executed appropriately, the reported rate constants were consistent, and independent of which protein was immobilized and which biosensor was used.

Original languageEnglish (US)
Pages (from-to)194-216
Number of pages23
JournalAnalytical Biochemistry
Volume386
Issue number2
DOIs
StatePublished - Mar 15 2009

Keywords

  • Biacore
  • Kinetics
  • Optical biosensor
  • Surface plasmon resonance

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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