Determining kinetics and affinities of protein interactions using a parallel real-time label-free biosensor, the Octet

被引:295
作者
Abdiche, Yasmina [1 ]
Malashock, Dan [1 ]
Pinkerton, Alanna [1 ]
Pons, Jaurne [1 ]
机构
[1] Pfizer Inc, Rinat Lab, San Francisco, CA 94080 USA
关键词
biacore; octet; ProteOn; protein/protein interaction; affinity;
D O I
10.1016/j.ab.2008.03.035
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
ForteBio's Octet optical biosensor harnesses biolayer interferometry to detect and quantify molecular interactions using disposable fiber-optic biosensors that address samples from an open shaking microplate without any microfluidics. We recruited a monoclonal antibody against a panel of peptides to compare the Octet directly with Biacore's well-established 3000 platform and Bio-Rad's recently launched ProteOn XPR36 array system, which use surface plasmon resonance (SPR) to detect the binding of one analyte over four surfaces and six analytes over six surfaces, respectively. A sink method was used to prevent analyte from rebinding the ligand-coated Octet tips and enabled us to extract accurate kinetic rate constants, as judged by their close agreement with those determined by SPR. Although the Octet is not sensitive enough to detect the binding of small molecules directly, it can access their affinities indirectly via solution competition experiments. We conducted similar experiments on the SPR instruments to validate these measurements. The Octet is emerging as a versatile complement to other more sophisticated biosensors, and the ProteOn provides high-quality data near the sensitivity of Biacore but in a more multiplexed format. Our results provide a benchmark for assessing the performance of the above-mentioned sensors. (c) 2008 Elsevier Inc. All rights reserved.
引用
收藏
页码:209 / 217
页数:9
相关论文
共 10 条
[1]   Pain pharmacology in migraine: focus on CGRP and CGRP receptors [J].
Benemei, S. ;
Nicoletti, P. ;
Capone, J. A. ;
Geppetti, P. .
NEUROLOGICAL SCIENCES, 2007, 28 (Suppl 2) :S89-S93
[2]   Exploring "one-shot" kinetics and small molecule analysis using the ProteOn XPR36 array biosensor [J].
Bravman, Tsafrir ;
Bronner, Vered ;
Lavie, Kobi ;
Notcovich, Ariel ;
Papalia, Giuseppe A. ;
Myszka, David G. .
ANALYTICAL BIOCHEMISTRY, 2006, 358 (02) :281-288
[3]   Kinetic exclusion assay technology: Characterization of molecular interactions [J].
Darling, RJ ;
Brault, PA .
ASSAY AND DRUG DEVELOPMENT TECHNOLOGIES, 2004, 2 (06) :647-657
[4]   Analyzing a kinetic titration series using affinity biosensors [J].
Karlsson, R ;
Katsamba, PS ;
Nordin, H ;
Pol, E ;
Myszka, DG .
ANALYTICAL BIOCHEMISTRY, 2006, 349 (01) :136-147
[5]  
Lowe PA, 1998, J MOL RECOGNIT, V11, P194, DOI 10.1002/(SICI)1099-1352(199812)11:1/6<194::AID-JMR422>3.0.CO
[6]  
2-T
[7]   Measuring long association phases using Biacore [J].
Navratilova, I ;
Eisenstien, E ;
Myszka, DG .
ANALYTICAL BIOCHEMISTRY, 2005, 344 (02) :295-297
[8]   Extracting kinetic rate constants from surface plasmon resonance array systems [J].
Rich, Rebecca L. ;
Cannon, Michelle J. ;
Jenkins, Jerry ;
Pandian, Prabhakar ;
Sundaram, Shankar ;
Magyar, Rachelle ;
Brockman, Jennifer ;
Lambert, Jeremy ;
Myszka, David G. .
ANALYTICAL BIOCHEMISTRY, 2008, 373 (01) :112-120
[9]   Higher-throughput, label-free, real-time molecular interaction analysis [J].
Rich, Rebecca L. ;
Myszka, David G. .
ANALYTICAL BIOCHEMISTRY, 2007, 361 (01) :1-6
[10]   MONOCLONAL-ANTIBODY TO RAT ALPHA-CGRP - PRODUCTION, CHARACTERIZATION, AND INVIVO IMMUNONEUTRALIZATION ACTIVITY [J].
WONG, HC ;
TACHE, Y ;
LLOYD, KCK ;
YANG, H ;
STERNINI, C ;
HOLZER, P ;
WALSH, JH .
HYBRIDOMA, 1993, 12 (01) :93-106