Kulim Kht SPR BioNavis reflectometer for liquids

Surface plasmon resonance instrument as a refractometer for liquids and ultrathin films

A surface plasmon resonance (SPR) setup in Kretschmann configuration is being utilized as a refractometer for both liquids as well as ultrathin films. The SPR signal detection technology used is based on a goniometer approach providing a wide angular scan range which facilitates highly accurate liquid and gas phase measurements.

Attention was paid to improve sample handling and preparation. In order to avoid cross-contamination between measurements an easily removable and exchangeable molded PDMS flow cell was used during the measurements. By careful choice of components for liquid handling the dead volume of the system could be reduced down to some microliters.

The angular change and thus the refractive index for sucrose, ethylene glycol (EG) and ethanol solutions with different concentrations, the thickness and refractive index of deposited Langmuir-Blodgett (LB) films, and the interaction kinetics between a biotin containing self-assembled monolayer (SAM) and streptavidin were determined. The measured refractive indices of sucrose, EG and ethanol solutions corresponded well with literature values. LB films were characterized by measuring the complete SPR curve in an angular scan range from 40 to 78[degree sign]. A two-color SPR approach combined with two-media measurements was successfully employed for simultaneous and unambiguous determination of both refractive index and thickness of stearic acid monolayers. The thickness obtained for the stearic acid monolayer was 2.66 nm, and the refractive indices at 635 and 670 nm were 1.5800 and 1.4138, respectively. The developed sensor-plate holder enabled functionalization of the SPR gold chip outside the instrument, therefore making the sample handling more flexible. The affinity constant obtained for the streptavidin-biotin interaction was 1.01 x 10-8 M. The total angle SPR method used in this study clearly shows its potential to be used as a refractometer for both liquids and ultrathin films, as well as for traditional liquid phase biomolecular kinetic studies.

"BioNavis' SPR Navi system allows unique configurations for multiple concurrent research projects"

Prof. Orlando Rojas
North Carolina State University in Raleigh
United States

Kulim Kht SPR BioNavis CMC

Modification of cellulose films by adsorption of CMC and chitosan for controlled attachment of biomolecules

The adsorption of human immunoglobulin G (hIgG) and bovine serum albumin (BSA) on cellulose supports were investigated. The dynamics and extent of related adsorption processes were monitored by surface plasmon resonance (SPR) and quartz crystal microbalance with dissipation monitoring (QCM-D). Amine groups were installed on the cellulose substrate by adsorption of chitosan from aqueous solution, which allowed for hIgG to physisorb from acid media and produced a functionalized substrate with high surface density (10 mg/m2). hIgG adsorption from neutral and alkaline conditions was found to yield lower adsorbed amounts. The installation of the carboxyl groups on cellulose substrate via carboxymethylated cellulose (CMC) adsorption from aqueous solution enhanced the physisorption of hIgG at acidic (adsorbed amount of 5.6 mg/m2) and neutral conditions. hIgG adsorption from alkaline conditions reduced the surface density. BSA was used to examine protein attachment on cellulose after modification with chitosan or carboxymethyl cellulose. At the isoelectric point of BSA (pI 5) both of the surface modifications enhanced the adsorption of this protein, when compared to that on unmodified cellulose (a two-fold increase from 1.7 to 3.5 mg/m2). At pH 4 the electrostatic interactions favored the adsorption of BSA on the CMC-modified cellulose, revealing the affinity of the system and the possibility of tailoring biomolecule binding and regeneration by choice of the surface modifier and pH of the medium.

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