Methods for site controlled coupling to carboxymethyldextran surfaces in surface plasmon resonance sensors

Abstract In the present study, optimized methods for in-situ ligand immobilization to carboxymethylated dextran coated gold surfaces for use in surface plasmon resonance (SPR) based biosensor devices were developed. Immobilization methods based on selective thiol reactions, either via thiols on the sensor surface or via thiols introduced on the ligand, were shown to give high yields of functionally active material. Methods are also described for linking aldehyde containing ligands to hydrazide modified surfaces and for coupling based on the avidin/biotin concept. The importance of blocking residual active groups (for thiol coupling) and reduction of acid labile hydrazone bonds formed in the aldehyde coupling was demonstrated. The methods were found to work efficiently in combination with conditions suitable for electrostatic preconcentration of ligands to be coupled, a concept we earlier developed for coupling of amine containing ligands (Lofas & Johnsson, 1990).

[1]  B. Persson,et al.  Quantitative determination of surface concentration of protein with surface plasmon resonance using radiolabeled proteins , 1991 .

[2]  G. Rechnitz.  The development of biocatalytic membrane electrodes , 1986 .

[3]  D. Christensen,et al.  Comparison of site-specific coupling chemistry for antibody immobilization on different solid supports , 1991 .

[4]  G. Whitesides,et al.  SELECTIVE REDUCTION OF DISULFIDES BY TRIS(2-CARBOXYETHYL) PHOSPHINE , 1991 .

[5]  P. Nilsson,et al.  Real-time monitoring of DNA manipulations using biosensor technology. , 1995, Analytical biochemistry.

[6]  R. Karlsson,et al.  Real-time biospecific interaction analysis using surface plasmon resonance and a sensor chip technology. , 1991, BioTechniques.

[7]  J Carlsson,et al.  Protein thiolation and reversible protein-protein conjugation. N-Succinimidyl 3-(2-pyridyldithio)propionate, a new heterobifunctional reagent. , 1978, The Biochemical journal.

[8]  A. Niveleau,et al.  The preparation of latex particles with covalently bound polyamines, IgG and measles agglutinins and their use in visual agglutination tests. , 1978, Journal of immunological methods.

[9]  C. Urbaniczky,et al.  Integrated fluid handling system for biomolecular interaction analysis. , 1991, Analytical chemistry.

[10]  C. Han,et al.  The conformation of γ-globulin adsorbed on polystyrene latices determined by quasielastic light scattering , 1978 .

[11]  Bo Johnsson,et al.  Comparison of methods for immobilization to carboxymethyl dextran sensor surfaces by analysis of the specific activity of monoclonal antibodies , 1995, Journal of molecular recognition : JMR.

[12]  M. Shipton,et al.  A reporter group delivery system with both absolute and selective specificity for thiol groups and an improved fluorescent probe containing the 7-nitrobenzo-2-oxa-1,3-diazole moiety. , 1975, The Biochemical journal.

[13]  S. Loefas,et al.  Immobilization of proteins to a carboxymethyldextran-modified gold surface for biospecific interaction analysis in surface plasmon resonance sensors. , 1991, Analytical biochemistry.

[14]  O'Shannessy Dj,et al.  Specific conjugation reactions of the oligosaccharide moieties of immunoglobulins. , 1985 .

[15]  D. O'Shannessy,et al.  Immobilization chemistries suitable for use in the BIAcore surface plasmon resonance detector. , 1992, Analytical biochemistry.

[16]  J. Butler,et al.  The physical and functional behavior of capture antibodies adsorbed on polystyrene. , 1992, Journal of immunological methods.

[17]  M. Wilchek,et al.  The avidin-biotin complex in bioanalytical applications. , 1988, Analytical biochemistry.

[18]  Bo Johnsson,et al.  A novel hydrogel matrix on gold surfaces in surface plasmon resonance sensors for fast and efficient covalent immobilization of ligands , 1990 .

[19]  Rolf Schmid,et al.  Biosensors: Applications in Medicine, Environmental Protection, and Process Control , 1989 .

[20]  S. Loefas,et al.  Dextran modified gold surfaces for surface plasmon resonance sensors: immunoreactivity of immobilized antibodies and antibody-surface interaction studies , 1993 .