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Published March 15, 2009 | Accepted Version
Journal Article Open

A global benchmark study using affinity-based biosensors

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


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.

Additional Information

© 2009 Elsevier B.V. Received 3 October 2008. Available online 27 November 2008. We thank KaloBios Pharmaceuticals for providing the purified Fab and GST–Ag, Biacore/GE Healthcare for providing sensor chips to help develop this model system, and Bio-Rad Laboratories for shipping all of the sample sets worldwide.

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Accepted Version - nihms500790.pdf


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