Determining kinetics and affinities of protein interactions using a parallel real-time label-free biosensor, the Octet.
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.
Abdiche
Yasmina
Y
Rinat Laboratories, Pfizer Inc., South San Francisco, CA 94080, USA. yasmina.abdiche@pfizer.com
Malashock
Dan
D
Pinkerton
Alanna
A
Pons
Jaume
J
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000378:metabolism; D019985:Benchmarking; D015374:Biosensing Techniques / Q000295:instrumentation / Q000379:methods*; D015740:Calcitonin Gene-Related Peptide / Q000276:immunology / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D051379:Mice; D011485:Protein Binding; D051381:Rats; D012680:Sensitivity and Specificity; D013194:Staining and Labeling; D020349:Surface Plasmon Resonance; D013997:Time Factors
D000911:Antibodies, Monoclonal; D015740:Calcitonin Gene-Related Peptide