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AN#170 Profiling of G protein-coupled receptor (GPCR) stimulation by small compounds in live cells
G protein-coupled receptors (GPCRs) are a large family of transmembrane proteins which are involved in many physiological and pathological processes. Extracellular stimuli result in cellular responses, which lead to ordered and dynamic redistribution of intracellular contents. Cellular responses are receptor-type specific. Accordingly, GPCRs are divided into four subclasses, Gi/o, Gq/11, Gs, and G12/13. Real-time label-free…
AN#169 Diagnostic method for serological testing of Covid-19 antibodies
Considering the indisputable need for Covid-19 related diagnostic tools, we have developed a dedicated serological testing method based on MP-SPR technology. The combination of MP-SPR instrument and optimized sensor format functionalized with virus proteins enabled rapid detection of SARS-CoV-2 antigen-specific antibodies in human serum. The measurement accuracy has been proven with seropositive patient samples and validated with certified reference serum materials. As alternative…
AN#168 Regenerable avidin kit assays Part II: Biomolecule affinity and concentration studies
MP-SPR Navi™ Regenerable avidin kit is an excellent tool in development of MP-SPR based assays. Here, the specific binding affinity of lipid based molecules towards sensor-captured protein was analyzed. Also, concentration analysis of diagnostically relevant biomarkers were performed in human serum samples. Utilizing Regenerable avidin kit reduces time in assay development since there is no…
AN#167 In-situ capture of T-cells and analysis of membrane receptor affinity with biofunctionalized lipid sensor
Assessment of T-cell specificity to tumor associated antigens is crucial for the development of personalized immunotherapies against cancer. Multi-Parametric Surface Plasmon Resonance (MP-SPR) was applied for characterization of interaction of T-cell receptors (TCR) from tumor-specific CD8+ T-cells. Intact live cells were captured onto biomimetic surfaces composed of artificial cell membranes functionalized with peptide-major histocompatibility complexes…
AN#166 Regenerable avidin kit assays Part I: Working principle
MP-SPR Navi™ Regenerable avidin kit is an excellent tool in development of MP-SPR based assays. The kit allows the user to conduct precise and high quality measurements in shorter time and convenient experimental set-up. Besides having the ligand available with biotin-conjugation, one does not need to optimize buffers and immobilization conditions. Finally, the cost effectiveness of SPR measurements improves since…
AN#165 Cellulose fiber-based yarn development for capturing estrogen residues from aqueous matrices with MP-SPR
In-situ preparation of chitosan and cyclodextrin-functionalized cellulose fiber yarns and capture of the synthetic estrogen hormone 17α-ethinyl estradiol (EE2) from water was investigated to address waste water purification. The thickness of functionalized chitosan and cyclodextrin layers were determined. The effectivity of the hormone capture was determined by determining the thickness of adsorbed EE2 layer.
AN#164 Affinity and kinetics of extracellular vesicles – protein interaction
Extracellular vesicles (EVs) are extensively studied as potential therapeutics, drug delivery systems and for non-invasive diagnostics of diseases, such as cancer. Early detection of ovarian cancer was explored using Multi-Parametric Surface Plasmon Resonance (MP-SPR) instrument. Tumor EVs (SKOV-3) were loaded onto surface-immobilized LXY30 peptide. Affinity and kinetic constants of the interaction, bound mass and layer…
AN#163 Characterization of micrometer thick layers of spin-coated chitin
Multi-Parametric Surface Plasmon Resonance (MP-SPR) instruments can measure optical properties and address surface interactions from very thin films (down to Ångstroms) up to micrometer thick layers. The measurement of thicker sensor coatings is based on a SPR waveguide mode. Micrometer thick chitin layers were spin-coated onto gold (Au) and polystyrene (PS) substrates and subsequently assessed through MP-SPR waveguide signal. Thickness and refractive…
AN#162 Concentration analysis of nutritionally important proteins from milk
Concentrations of proteins were determined from milk and milk-based infant formula using Multi-Parametric Surface Plasmon Resonance (MP-SPR) instrument. α-Lactalbumin (ALAB) is a nutritionally important protein for infants, and immunoglobulin G (IgG) is an important part of the secondary immune response. Concentrations of unknown samples were determined based on calibration curves. The assay was a direct…
AN#161 Polymerization kinetics and morphology changes of thermo-responsive polymer
Multi-Parametric Surface Plasmon Resonance (MP-SPR) measures surface changes in real-time. High sensitivity and minute volumes make the measurement method excellent for thin film characterization providing layer thickness and refractive index. Thermo-responsive poly(N-isopropyl-acrylamide) (PNIPAM) polymerization kinetics was measured during atom transfer radical polymerization (ATRP) in real-time using MP-SPR. Polymer growth was linear with a rate of ~20 nm/min up to 100 nm after which chain…