Preparation and use of a reactive solid support surface

a solid support surface and reactive technology, applied in the field of solid support surface preparation, to achieve the effect of high binding agent level and effective resistance to non-specific binding

Inactive Publication Date: 2006-05-04
GE HEALTHCARE BIO SCI CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0013] The above and other objects and advantages are provided by a method for preparing a solid support surface with immobilized binding agent(s) and a protein resistant compound, which method basically is characterized in that the solid support surface has a hydrogel coating, and that (i) a protein resistant compound, such as, e.g., poly(ethylene glycol) or a derivative thereof, and (ii) a binding agent(s) are coupled to the hydrogel to be co-immobilized thereon. In this way, it is possible to provide a solid support surface, such as a sensing surface, which has a high level of binding agent, e.g., analyte-specific ligand, simultaneously with a sufficient level of a protein resistant compound to effectively resist non-specific binding.

Problems solved by technology

In, for example, immunogenicity studies where antibodies in serum samples are analyzed, problems in most cases arise due to non-specific binding from non-analyte species in the serum, this non-specific binding often being greater than the specific binding of the target antibody.

Method used

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  • Preparation and use of a reactive solid support surface
  • Preparation and use of a reactive solid support surface

Examples

Experimental program
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Effect test

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Instrumentation

[0078] A Biacore® 3000 (Biacore AB, Uppsala, Sweden) was used. This instrument, which is based on surface plasmon resonance (SPR) detection at a gold surface, uses a micro-fluidic system for passing samples and running buffer through four individually detected flow cells, designated Fc1 to Fc4, one by one or in series. As sensor chip was used Series CM5 (Biacore AB, Uppsala, Sweden) which has a gold-coated (about 50 nm) surface with a covalently linked hydrogel matrix (about 100 nm) of carboxymethyl-modified dextran polymer. As running buffer was used HBS-EP (Biacore AB). Unless indicated otherwise, the flow rate was 5 μl / min. The output from the instrument is a “sensorgram” which is a plot of detector response (measured in “resonance units”, RU) as a function of time. An increase of 1000 RU corresponds to an increase of mass on the sensor surface of approximately 1 ng / mm2.

Rat Sera

[0079] In the experiments below were used sera from rats (Wistar F) which had and h...

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Abstract

A method of preparing a protein-resistant reactive solid support surface is disclosed. The method comprises the steps of providing a solid support having a hydrogel coating with a plurality of binding elements, coupling a protein resistant compound to the hydrogel via a first fraction of the binding elements, and coupling at least one binding agent to the hydrogel via a second fraction of the binding elements, whereby the protein resistant compound and the at least one binding agent are co-immobilized to the hydrogel. Also the use of the reactive surface in analysis, such as immunogenicity assays, is disclosed.

Description

CROSS-REFERENCE TO RELATED APPLICATION [0001] This application claims the benefit under 35 U.S.C. § 119(e) of U.S. Provisional Patent Application No. 60 / 618,655 filed Oct. 13, 2004; and also claims priority to Swedish Application No. 0402476-6 filed Oct. 13, 2004; both of these applications are incorporated herein by reference in their entireties.BACKGROUND OF THE INVENTION [0002] 1. Field of the Invention [0003] The present invention relates to a method for preparing a solid support surface having binding agents, such as analyte-specific ligands, immobilized thereto, and more particularly to such a surface which resists non-specific binding. The invention also relates to the use of the prepared solid support surface in analysis, such as immunogenicity assays, and to a protein-resistant solid support surface for coupling of binding agents thereto. [0004] 2. Description of the Related Art [0005] A variety of analytical techniques are used to characterize interactions between molecule...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): G01N33/53B05D3/00G01N
CPCB82Y30/00G01N33/548G01N2800/52Y10S436/809
Inventor JARHEDE, TANJAKJELLIN, PERLARSSON, ANITASJOBOM, HANS
Owner GE HEALTHCARE BIO SCI CORP
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