Label-Free Free Solution Single Protein-Small Molecule Binding Kinetics: An Optical Tweezer Approach

2015 ◽  
Author(s):  
Ahmed A. Al Balushi ◽  
Reuven Gordon
Keyword(s):  
Small Molecule ◽  
Optical Tweezer ◽  
Binding Kinetics ◽  
Free Solution ◽  
Label Free ◽  
Single Protein

scholarly journals Highly Multiplexed Label-Free Imaging Sensor for Accurate Quantification of Small-Molecule Binding Kinetics

ACS Omega ◽  
2020 ◽  
Vol 5 (39) ◽  
pp. 25358-25364
Author(s):  
Elisa Chiodi ◽  
Allison M. Marn ◽  
Matthew T. Geib ◽  
Fulya Ekiz Kanik ◽  
John Rejman ◽  
...  
Keyword(s):  
Small Molecule ◽  
Binding Kinetics ◽  
Label Free ◽  
Imaging Sensor ◽  
Accurate Quantification

scholarly journals Highly Multiplexed Label-Free Imaging Sensor for Accurate Quantification of Small-Molecule Binding Kinetics

Proceedings ◽  
2020 ◽  
Vol 60 (1) ◽  
pp. 37
Author(s):  
Elisa Chiodi ◽  
Allison M. Marn ◽  
Matthew T. Geib ◽  
Fulya Ekiz Kanik ◽  
John Rejman ◽  
...  
Keyword(s):  
Small Molecule ◽  
Limit Of Detection ◽  
Signal To Noise Ratio ◽  
Fumonisin B1 ◽  
Biomass Accumulation ◽  
Binding Kinetics ◽  
Label Free ◽  
Common Path ◽  
Label Free Detection ◽  
Imaging Sensor

Investigating the binding kinetics of small molecule analytes to larger ligands, such as proteins and antibodies, is a compelling task for the field of drug and biomarker development, as well as the food industry and agro-biotechnology. Here, we improve the limit of detection of the Interferometric Reflectance Imaging Sensor (IRIS), a label-free, highly multiplexed biosensor, to perform real-time affinity measurement of small molecules binding to immobilized antibodies in a microarray format. As the analytes bind to the surface probes, the biomass accumulation on the surface is quantified by measuring the optical reflectance from the layered Si/SiO2 chip through the solution, in a common-path interferometer configuration. As a proof of concept, label-free detection of biotin molecules binding to immobilized streptavidin probes is performed, achieving 1 pg/mm2 sensitivity through signal averaging in a shot noise limited operation. Furthermore, we apply the optimized sensor to the screening of a 20-multiplexed antibody chip (MW~150 kDa ligands) against Fumonisin B1 (MW = 721.8 Da), one of the most prevalent mycotoxins found in many cereal grains such as corn and wheat. The simultaneously recorded binding curves of the toxin to the multiplexed sensor yield a signal-to-noise ratio of ≈8 when noise reduction methods of spatial and temporal averaging are utilized.

Pre-equilibration kinetic size-exclusion chromatography with mass spectrometry detection (peKSEC-MS) for label-free solution-based kinetic analysis of protein–small molecule interactions

The Analyst ◽  
2015 ◽  
Vol 140 (4) ◽  
pp. 990-994 ◽  
Author(s):  
Jiayin Bao ◽  
Svetlana M. Krylova ◽  
Leonid T. Cherney ◽  
J. C. Yves Le Blanc ◽  
Patrick Pribil ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Kinetic Analysis ◽  
Small Molecule ◽  
Size Exclusion ◽  
Free Solution ◽  
Label Free ◽  
Mass Spectrometry Detection ◽  
Reversible Binding ◽  
Exclusion Chromatography ◽  
Molecule Interactions

Label-free solution-based kinetic analysis of reversible binding between protein and small molecule.

scholarly journals Critical angle reflection imaging for quantification of molecular interactions on glass surface

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Guangzhong Ma ◽  
Runli Liang ◽  
Zijian Wan ◽  
Shaopeng Wang
Keyword(s):  
Refractive Index ◽  
Small Molecule ◽  
Molecular Interactions ◽  
Glass Surface ◽  
Critical Angle ◽  
Dynamic Range ◽  
Refractive Index Change ◽  
Label Free ◽  
Index Change ◽  
Sensing Range

AbstractQuantification of molecular interactions on a surface is typically achieved via label-free techniques such as surface plasmon resonance (SPR). The sensitivity of SPR originates from the characteristic that the SPR angle is sensitive to the surface refractive index change. Analogously, in another interfacial optical phenomenon, total internal reflection, the critical angle is also refractive index dependent. Therefore, surface refractive index change can also be quantified by measuring the reflectivity near the critical angle. Based on this concept, we develop a method called critical angle reflection (CAR) imaging to quantify molecular interactions on glass surface. CAR imaging can be performed on SPR imaging setups. Through a side-by-side comparison, we show that CAR is capable of most molecular interaction measurements that SPR performs, including proteins, nucleic acids and cell-based detections. In addition, we show that CAR can detect small molecule bindings and intracellular signals beyond SPR sensing range. CAR exhibits several distinct characteristics, including tunable sensitivity and dynamic range, deeper vertical sensing range, fluorescence compatibility, broader wavelength and polarization of light selection, and glass surface chemistry. We anticipate CAR can expand SPR′s capability in small molecule detection, whole cell-based detection, simultaneous fluorescence imaging, and broader conjugation chemistry.

Label-free detection of small-molecule binding to a GPCR in the membrane environment

2015 ◽  
Vol 1854 (8) ◽  
pp. 979-986 ◽  
Author(s):  
Roland G. Heym ◽  
Wilfried B. Hornberger ◽  
Viktor Lakics ◽  
Georg C. Terstappen
Keyword(s):  
Small Molecule ◽  
Label Free ◽  
Label Free Detection
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