Surface Plasmon Resonance (SPR) is a widely-used label-free technique to characterize a variety of molecular interactions. SPR is an optical phenomenon that is sensitive to changes in the dielectric properties of the medium close to a metal surface. Specifically, the resonance condition is affected by changes in refractive index occurring up to 300 nm above the metal surface (Au) and thus by the material absorbed onto the metal film. Therefore, the SPR signal is a measure of the total mass concentration at the gold sensor chip surface.
Typically, a mobile molecule (analyte) is injected across an immobilized binding partner (ligand) and as the analyte binds, this mass accumulation on the sensor surface leads to an increase in refractive index, and the result is plotted as response versus time. SPR is commonly utilized by researchers to determine association/dissociation rates, affinities and thermodynamics of biomolecular interactions.
Traditionally, the interactions under study with SPR include those occurring with and between the major classes of biological macromolecules along with those involving small molecules and drugs. These classic experiments have been primarily carried with just purified samples. Reichert’s SPR systems implement a very robust fluidics arrangement that can accommodate a wide variety of sample compositions including crude samples such as lysates, whole cells and serum. In addition, Reichert’s systems are housed in an open architecture that easily allows coupling to other analytical techniques and instruments. Along with excelling at traditional biomolecular interactions, Reichert’s systems pave the way for new avenues of investigation involving crude samples and whole cells along with the ability to couple SPR to other techniques. This presentation will focus on the SPR technique and provide examples of unique applications with cells along with the possibility of interfacing SPR with other analytical methods.
The webinar demonstrates the use of SPR for measurement of cell adhesion interactions in biomedical applications.
Various biotechnology applications benefit from studies of cell-substrate interactions. A number of laboratories in the fields of biomedicine, biotechnology, and biophysics are keenly pursuing the nature of specific, receptor-ligand type interactions, and non-specific physical adhesive interactions.
The Reichert SR7500DC dual channel system, a surface plasmon resonance (SPR) instrument with unmatched sensitivity and high-quality binding data for interaction analysis, is ideally suited for these applications. Researchers can use a large-tubing diameter that allows for the perfusion of cells, and the instrument’s open-design architecture accommodates ready manipulation of the SPR substrate.
Sponsored by Reichert Technologies Life Sciences, this free educational webinar, “Use of Surface Plasmon Resonance for Probing Cell-Matrix Interactions,” demonstrates the use of SPR for the measurement of cell adhesion interactions in varied biomedical applications, and presents two examples in which the Reichert SPR system has been used for studying cells. The first example discusses human white blood cell (HL-60) adhesion/capture by the endothelial cell adhesion molecule P-selectin. Several antibody- and recombinant protein-based controls are used to demonstrate the application of SPR for human vascular-biology research. The second example discusses binding interactions between endothelial cells and two different extracellular matrix proteins, Collagen 1 and Matrigel.
The speaker is research assistant Michael Hill. Michael who completed his PhD in biomedical engineering at the University of Buffalo this year, utilizing the SPR data in his thesis. Michael’s advisor, Debanjan Sarkar, PhD, is in the laboratory of biomaterials and regenerative therapeutics.
The free webinar was hosted by LabRoots, and was presented on September 20, 2016.
The use of surface plasmon resonance to characterize protein:protein interactions is well established and has been the method of choice for determination of antibody affinity for the last 20 years. Developments such as phage display have led to rapid production of numerous antibodies in a high-throughput environment that typically requires affinity analysis to select the proper candidates for further development. Moreover, since most display libraries are limited in terms of the number of sequences in the libraries (typically 1x109 to 1x1010), a frequent result is that initial candidates for development are not of sufficiently high affinity, necessitating affinity improvement strategies. In this presentation, we will describe our studies to demonstrate that the SR7000DC can be used to improve the efficiency of phage antibody isolation and characterization and to evaluate the potential for optimizing selection of high affinity variants. The idea of using an SPR instrument to function as both a lab-on a chip to permit semi-automated manipulation of phage preparations as well as providing the SPR signal as a window on the binding and enrichment of high affinity variants of scFvs or Fabs (or other scaffolds like camelid VHH and fibronectin domains) is appealing, and could lead to a significant competitive advantage in the expanding market for therapeutic antibodies.
Surface plasmon resonance (SPR) is a sensitive, label-free technique that detects mass changes due to biomolecular interactions on a surface. This versatile method has been used to evaluate the specificity and binding kinetics of molecules, ranging from short oligonucleotides to whole cells. In this webinar, we will discuss the use of SPR to investigate interactions between aptamers and their targets, as well as the triggered formation of DNA polymers and polyvalent aptamers. In addition, the use of oligonucleotides for the release of protein drugs will be discussed.