Surface Plasmon Resonance (SPR) has been used for measurements of molecular interactions for three decades. The SPR technology allows evaluation of affinity and kinetics of the molecular binding reaction without the burdens of labeling as the technique is label-free.
Molecular interaction measurements can be performed in diverse liquids, including complex liquids such as serum, saliva or organic solvents.
The unique PureKinetics™ feature together with large working range make MP-SPR an essential tool for new challenges posed by the shift from synthetic to biopharmaceutic drugs. From antibody characterization through drug uptake routes, controlled drug release strategies, small molecule measurements, nanoparticle targeting up to drug internalization by living cells, MP-SPR helps you to get ahead of competition.
MP-SPR is a sensitive real-time label-free method that not only provides affinity but also kinetic information on molecular interactions. Even molecules of equal affinity can have very different kinetic profiles making kinetics essential selection parameters especially in drug discovery.
To determine affinity and kinetic constants, the MP-SPR data are plotted against time as sensograms. It is possible to select from a number of parameters to be plotted, including Peak Minimum Angle. Such sensograms are fitted with binding models in data analysis software TraceDrawer™ for MP-SPR Navi™
Anatomy of a kinetics measurement
- 0-2 min: running buffer is passed over the sensor surface with one of the interaction partners (ligand) immobilized on the sensor surface.
- 2-6 min: The second interaction partner (analyte) is run over the surface. As the analytes attaches on the surface, we can observe an association phase.
- 6-8 min: Running buffer is subsequently run over the surface with attached ligand-analyte. The buffer will remove (dissociate) any unbound analyte.
- 8-10 min: Finally, regeneration buffer is run over the surface to regain clean sensor surface.
- 10-12 min: Running buffer is passed over the sensor. Next sample injection can follow. (Reuse of sensors is possible for most cases).
Affinity and kinetics data can be collected from proteins, peptides, nucleotides, drug molecules, viruses, nanoparticles, living cells, bacteria, biomaterials and more.
Several methods for ligand capturing are available such as direct coupling, affinity capture, or membrane anchoring, depending on the molecule attached. To find out the best strategy for your sample analysis, contact our scientific team.
MP-SPR is a sensitive platform to determine antibody and fragment binding on antigen. Label-free interactions are measured in real-time, revealing affinity and kinetics of the binding. MP-SPR instruments can also handle target molecules incorporated in liposomes (membrane extract) and living cells, enabling interaction measurements in natural-like environment.
Interaction measurements can be performed in diverse fluids including 100% serum, plasma or cell growth medium. With PureKinetics™, bulk effect can be distinguished from binding and thus high precision results are obtained. Fitting SPR curves with LayerSolver™ provides insight into the conformation and orientation of the antibodies via thickness assessment of the adsorbed layers.
Reasons to choose MP-SPR for antibody characterization:
- Label-free affinity and kinetics
- Low total cost of ownership
- Conformation changes
- Crude samples (100% serum)
- Biopharmaceuticals even from cell medium
- Easy ex-situ preparation of own sensor surfaces
- Robust fluidics to handle harsh chemicals
With MP-SPR:
- Determine an antibody’s affinity and kinetics to antigens
- Select the best antibody for successful immunoassays
- Measure antibody binding on receptor in lipid environment
- Characterize which cell signaling cascade an antibody activates in living cells
- Identify the monoclonal antibody with the highest affinity
- See the real binding without bulk effect
Not yet convinced? See selected Application Notes to see how MP-SPR works:
- AN#169 Diagnostic method for serological testing of Covid-19 antibodies
- AN#168 Regenerable avidin kit assays Part II: Biomolecule affinity and concentration studies
- AN#166 Regenerable avidin kit assays Part I: Working principle, antibody – antigen interaction
- AN#162 Concentration analysis of nutritionally important proteins from milk
- AN#147 Analyzing dissociation kinetics of IgG from protein A using MP-SPR and PureKinetics™
Reasons to choose MP-SPR for small molecule interactions:
- Direct measurements of small molecules
- High quality data with PureKinetics™
- Low total cost of ownership
- Easy shift from targeting to internalization studies
- Answers to new challenges in biopharmaceutical development and manufacturing
With MP-SPR:
- Determine the affinity and kinetics of small molecular weight drugs to its target protein
- Select the molecules that binds best to target receptor
- Measure the release rate of small molecular weight drug from delivery materials
- Characterize the real binding without bulk effect
Not yet convinced? See selected Application Notes to see how MP-SPR works:
Reasons to choose MP-SPR for drug delivery studies:
- MP-SPR’s angular range allows to measure large shifts in SPR caused by nanoparticles
- MP-SPR follows a nanocarrier’s targeting and internalization in the same measurement
- No more clogged flow-cells or tubings: our customizable MP-SPR instruments allow for easy exchange of fluidics by the user
- From static to dynamic measurements of nanoparticles: MP-SPR measures affinity and kinetics of nanoparticle adsorption
- From SEM to MP-SPR and back to SEM: easy removal of substrates for ex-situ modification or measurements
With MP-SPR:
- Determine the best nanoparticle for drug delivery
- Characterize the absorption route of a drug nanocarrier
- Elucidate how nanoparticles interact with lipid bilayers
- Measure the release rate of drug from carrier materials
- Identify how a nanoparticle or virus enters the cell
Controlled drug release formulations are extensively studied to enhance or extend drug absorption. MP-SPR provides drug release rates and enables in vitro comparison of drug release formulations.
Reasons to choose MP-SPR for controlled drug release:
- Quantify drug release up to micron thick samples
- Measure real-time and label-free
- Get rid of bulk effect
- Understand structural changes
Not yet convinced? See selected Application Notes to see how MP-SPR works:
- AN#152 Drug delivery nanocarrier studies using MP-SPR