Biomedical subjects

Stefan Löfås

Publications and source records attributed to Stefan Löfås.

5 recordsLinked to original sources

PubMed2006

Surface plasmon optical studies of carboxymethyl dextran brushes versus networks.

The conformational changes of carboxymethyl dextran (CMD) substrates induced by variations in pH and ionic strength were studied using surface plasmon field-enhanced fluorescence spectroscopy (SPFS). A typical response was an increase in swelling upon increasing the pH and decreasing the ionic strength. Furthermore, the effects of the surface charge and cross-link density of the CMD on the degree of stimulus responses were investigated. The swelling/collapse ratio decreased with decreasing carboxyl group surface concentration and increasing cross-link density.

Dextrans↗

PubMed2004

Surface plasmon fluorescence immunoassay of free prostate-specific antigen in human plasma at the femtomolar level.

This study describes the development of a surface plasmon fluorescence spectroscopy (SPFS)-based sandwich immunoassay, for the detection of free prostate-specific antigen (f-PSA). The commercial CM5 chip (Biacore) was integrated into the SPFS-based assay making use of its three-dimensionally extended dextran architecture, which offers a large protein retention capacity and alleviates the metal-induced fluorescence quenching. The performance of the f-PSA assay was investigated in buffer and in human plasma, respectively. In the human plasma, the limit of detection of f-PSA was estimated to be approximately 80 fM for 40 min of contact time, which adequately meets clinical requirements.

Humans↗

PubMed2004

Attomolar sensitivity in bioassays based on surface plasmon fluorescence spectroscopy.

With the aid of a dextran matrix, the metal-induced fluorescence loss of bound fluorophores can be greatly reduced, and the distance dependence of the fluorescence yield could be largely convoluted. This is optimized for the limit of detection assessment of surface plasmon fluorescence spectroscopy. The model system was designed as a direct assay with mouse IgG covalently immobilized to the carboxymethyl dextran matrix of a CM5 sensor chip from Biacore. Time-resolved ultratrace detection of fluorophore (Alexa-Fluor 647)-labeled rabbit anti-mouse antibody down to 500 aM (10-18 M) was accomplished, corresponding to a binding rate of approximately 10 molecules mm-2 min-1.

Animals↗

PubMed2004

Optimizing the hit-to-lead process using SPR analysis.

Secondary screening and lead optimization, where a large number of "hit" compounds are refined to a viable set of "lead" drug candidates, are considered to be bottlenecks to the drug discovery process and are targets for streamlining. Surface plasmon resonance (SPR) is a nonlabel technology that can generate kinetic data on biomolecular interactions. This allows researchers to quantitate the binding characteristics of lead compounds with their targets in terms of affinity, specificity, and association/dissociation rates in parallel. The latest generation of SPR biosensors integrate the hit-to-lead process and generate a greater depth of information, providing answers that cannot be addressed by traditional end-point assays. This allows users to make more informed choices on the selection of candidate molecules prior to preclinical development. A number of studies have used SPR biosensors in secondary screening, lead optimization, quantitative structure-activity relationship analysis, and predictive adsorption, distribution, metabolism, excretion, and/or toxicity evaluations.

Biosensing Techniques↗

PubMed2002

Flow-mediated on-surface reconstitution of G-protein coupled receptors for applications in surface plasmon resonance biosensors.

To facilitate biosensor studies of G-protein coupled receptors (GPCR) and other membrane proteins, reliable methods for preparation of sensor surfaces with high protein density are required. We present here a method for the easy and rapid immobilization and reconstitution of GPCR on carboxylated dextran surfaces modified with long alkyl groups. Following amine coupling of the detergent-solubilized receptor, lipid/detergent-mixed micelles were adhered as they were injected over the immobilized surface, taking advantage of the integrated flow cells. The detergent was eluted in the subsequent buffer flow and the remaining lipid formed a bilayer on the chip surface. With this procedure, rhodopsin was functionally reconstituted in a lipid environment in approximately 1 min. This method can also be used for the easy formation of pure supported lipid bilayers for use in model membrane interaction studies.

Heterotrimeric GTP-Binding Proteins↗