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A global benchmark study using affinity-based biosensors |
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| Affiliation: | a Center for Biomolecular Interaction Analysis, School of Medicine, University of Utah, Salt Lake City, UT 84132, USA b KaloBios Pharmaceuticals, South San Francisco, CA 94080, USA c Schering-Plough Biopharma, Palo Alto, CA 94304, USA d Nomadics, Oklahoma City, OK 73104, USA e Biochemistry and Molecular Biophysics, California Institute of Technology, Pasadena, CA 91125, USA f Department of Biochemistry and Molecular Biophysics, Columbia University, New York, NY 10032, USA g Hartwell Center for Bioinformatics and Biotechnology, St. Jude Children’s Research Hospital, Memphis, TN 38105, USA h University of Zurich, CH-8057 Zurich, Switzerland i Molecular Probes/Invitrogen, Eugene, OR 97402, USA j University of Strasbourg, 67412 Illkirch, France k Merck KGaA, D-64293 Darmstadt, Germany l Abbott Bioresearch Center, Worcester, MA 01605, USA m Institute of Molecular and Cellular Biology, University of Strasbourg, 67084 Strasbourg, France n Reichert, Depew, NY 14043, USA o Momenta Pharmaceuticals, Cambridge, MA 02142, USA p XOMA (U.S.), Emeryville, CA 94608, USA q Department of Chemical and Biomolecular Engineering, National University of Singapore, Singapore 119260, Singapore r Department of Biology, University of Ljubljiana, 1000 Ljubljiana, Slovenia s Department of Biological Chemistry, Weizmann Institute of Science, 76100 Rehovot, Israel t Molecular Biotechnology Core Laboratory, Cleveland Clinic Foundation, Cleveland, OH 44195, USA u ThermoFisher Scientific, Rockford, IL 61101, USA v Biacore/GE Healthcare, SE-754 50 Uppsala, Sweden w AstraZeneca, Hayward, CA 94545, USA x Neurodegeneration Research Department, GlaxoSmithKline, Harlow CM19 5AW, UK y MedImmune, Gaithersburg, MD 20878, USA z Biacore/GE Healthcare, Piscataway, NJ 08854, USA aa Department of Biochemistry, Medical College of Wisconsin, Milwaukee, WI 53226, USA ab Alexion Pharmaceuticals, Cheshire, CT 06410, USA ac Applied Biosystems, Foster City, CA 94404, USA ad diaDexus, South San Francisco, CA 94080, USA ae Eli Lilly, Indianapolis, IN 46285, USA af Laboratory of Thrombosis and Haemostasis, Center for Applied Medical Research, University of Navarra, 31008 Pamplona, Spain ag EMD Lexigen Research Center, Billerica, MA 01821, USA ah Department of Cell Biology and Neuroscience, Istituto Superiore di Sanità, 00161 Rome, Italy ai Department of Structural Biology and Chemistry, Pasteur Institute, 75724 Paris, France aj Department of Chemistry, Georgia State University, Atlanta, GA 30303, USA ak Pfizer/Rinat Laboratories, South San Francisco, CA 94080, USA al PDL BioPharma, Fremont, CA 94555, USA am Department of Biochemistry, University of Texas Health Science Center at San Antonio, San Antonio, TX 78229, USA an Akubio, Cambridge CB4 0GJ, UK ao INSERM U869, Institut Europeen de Chimie et Biologie, University of Bordeaux, 33607 Pessac, France ap Wyeth Research, Cambridge, MA 02140, USA aq Department of Biochemistry and Molecular Biology, University of Southern Denmark, DK-5230 Odense M, Denmark ar NovoNordisk, 2820 Gentofte, Denmark as Södertörns University College, 14152 Huddinge, Sweden at Department of Biochemistry, Temple University, Philadelphia, PA 19140, USA au U.S. Food and Drug Administration, Bethesda, MD 20892, USA av Department of Biochemical Engineering, University College London, London WC1E 7JE, UK aw Merck, 00040 Rome, Italy ax Roche, Palo Alto, CA 94304, USA ay University of Twente, 7500AE Enschede, The Netherlands az Agensys, Santa Monica, CA 90404, USA ba Novartis, Emeryville, CA 94608, USA bb Massachusetts Institute of Technology, Cambridge, MA 02139, USA bc Northern Illinois University, DeKalb, IL 60115, USA bd Pasteur Institute of Montevideo, CP 11400 Montevideo, Uruguay be University of Manchester, Manchester M1 7ND, UK bf Department of Physiology, University of Maryland, Baltimore, MD 21201, USA bg Complex Carbohydrate Research Center, University of Georgia, Athens, GA 30602, USA bh Adnexus Therapeutics, Waltham, MA 02453, USA bi University of Tours, 37380 Nouzilly, France bj ForteBio, Menlo Park, CA 94025, USA bk University of Michigan, Ann Arbor, MI 48109, USA bl Human Genome Sciences, Rockville, MD 20850, USA bm Department of Chemical Sciences, University of Catania, 95125 Catania, Italy bn Montana State University, Bozeman, MT 59717, USA bo GKT School of Biomedical Sciences, London SE1 1UL, UK bp Organic and Pharmaceutical Chemistry Department, Complutense University, 28040 Madrid, Spain bq Bio-Rad Haifa, 32000 Haifa, Israel br Institute of Chemistry, Academia Sinica, Taipei 115, Taiwan bs University of Texas Medical School at Houston, Houston, TX 77030, USA bt Fred Hutchinson Cancer Research Center, Seattle, WA 98109, USA bu Division of Molecular Structure, National Institute of Medical Research (UK), London NW7 1AA, UK bv Centre for Translational and Chemical Biology, Institute of Structural and Molecular Biology, University of Edinburgh, Edinburgh EH9 3JR, UK bw Acceleron Pharma, Cambridge, MA 02139, USA bx National Research Council (Canada), Biotechnology Research Institute, Montreal, Que., Canada H4P 2R2 by CSIRO Health Sciences and Nutrition, Parkville, VIC 3052, Australia bz Battelle Biomedical Research Center, Columbus, OH 43201, USA ca Attana AB, SE-113 47 Stockholm, Sweden cb Corning, Corning, NY 14831, USA cc School of Life Sciences, Institute for Chemistry and Bioanalytics, University of Applied Sciences Northwestern Switzerland, CH-4132 Muttenz, Switzerland cd Bio-Rad, Hercules, CA 94547, USA ce Monsanto, Chesterfield, MO 63198, USA cf Genzyme, Cambridge, MA 02142, USA cg Saarland University, D-66041 Saarbrücken, Germany ch Institute of Neuroimmunology of SAS, 84510 Bratislava, Slovakia ci Bristol-Myers Squibb, Princeton, NJ 08543, USA cj Array Biopharma, Boulder, CO 80301, USA ck CEA, iBiTecs, Service d’Ingénierie Moléculaire des Protéines, 91191 Gif sur Yvette, France cl Department of Biochemistry, Masaryk University, 61137 Brno, Czech Republic cm University of Connecticut Health Center, Farmington, CT 06030, USA cn University of Southern California, Los Angeles, CA 90033, USA co GeminX Biotechnologies, Montreal, Que., Canada H2X 2H7 cp Hoffman-La Roche, Nutley, NJ 07110, USA cq IRBM, Pomezia, 00040 Rome, Italy cr Ludwig Institute for Cancer Research, Royal Melbourne Hospital, Parkville, Vic. 3050, Australia cs Regeneron, Tarrytown, NY 10591, USA ct University of Muenster, D-48149 Muenster, Germany cu Department of Molecular Biology, University of Salzburg, A-5020 Salzburg, Austria cv XanTec Bioanalytics, D-48149 Muenster, Germany cw Biosciences Division, Los Alamos National Laboratory, Los Alamos, NM 87545, USA cx Lumera, Bothell, WA 98011, USA cy Arizona State University, Tempe, AZ 85287, USA cz Dyax, Cambridge, MA 02139, USA da Lombardi Cancer Center, Georgetown University Medical Center, Washington, DC 20057, USA db Department of Biochemistry, University of Washington, Seattle, WA 98195, USA dc Zymogenetics, Seattle, WA 98102, USA dd Bio-Rad Canada, Edmonton, Alta., Canada T6R 2W6 de Bio-Rad Canada, Toronto, Ont., Canada |
| 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. |
| Keywords: | Biacore Kinetics Optical biosensor Surface plasmon resonance |
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