US 6,586,232 B2Granted

Substrate preparation for chemical-species-specific binding

AssigneeAgilent Technologies, Inc.

InventorsMay Tom-Moy, Carl Alan Myerholtz

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Abstract

A mass biosensor uses an intermediate avidin layer to facilitate binding of a biotinylated antibody to a measurement surface of the biosensor. The avidin layer can be added by the manufacturer of the biosensor, while the biotinylated layer can be added by the user. This two-phase method of chemically modifying the measurement surface significantly reduces the user time required to customize the measurement surface to render it capable of binding selected compounds. An organosilane coupling agent attached to the surface provides sites to which avidin is bound. Avidin acts as a universal receptor of biotinylated compounds with specific binding affinities. Biotinylated antibodies or other biotinylated compounds are added and bind to the immobilized avidin. Surface adsorption is reduced by washing the modified surface with biotin to block potential sites of weak bond formation, electrostatic and hydrophobic interactions.

Claims3

Dependent claims

A method for rendering a substrate surface specific for the binding of a selected substance, said method comprising the steps of:

Attaching to the surface of said substrate a layer of coupling agent which binds the layer to said substrate and which possesses binding sites for binding a ligand-binding substance thereto wherein said coupling agent is silyl compound of the formula:
RnSiX(4-n)
wherein R is a nonhydrolyzable organic radical containing a primary alcohol group, X is a hydrolyzable moiety selected from the group consisting of chloro, amino, alkoxy, and acyloxy, and n is an integer from 1 to 3;
attaching a layer of ligand-binding substance to said layer of coupling agent, said layer of ligand-binding substance having reciprocal binding sites available for binding said ligand-binding substance to said coupling agent as well as binding sites available for binding said ligand-binding substance to a ligand-bearing substance;
and
attaching a layer of ligand-bearing substance to said ligand-binding layer, said ligand-bearing substance having reciprocal binding sites available for binding said ligand-bearing substance to said ligand-binding substance as well as binding sites for binding of the selected substance;
and
washing the resulting structure with a blocking agent.

5 more elements

Dependencies are inferred from the claim text.

Facts

Application: 09/814,360Filed 21 Mar 2001 · division
Priority: 29 Sep 1988
Published: US 2002/0192718 A119 Dec 2002
Granted: 1 Jul 2003
Term ends: Not estimatedThe source records don’t support an estimate.

Classification11 codes

G01N 33/54373
Apparatus specially adapted for solid-phase testing: involving physiochemical end-point determination, e.g. wave-guides, FETS, gratings
G01N 29/022
Analysing fluids: fluid sensors based on microsensors, e.g. quartz crystal-microbalance [QCM], surface acoustic wave [SAW] devices, tuning forks,…
G01N 33/54353
With an insoluble carrier for immobilising immunochemicals: with ligand attached to the carrier via a chemical coupling agent
G01N 33/552
The carrier being inorganic: glass or silica

Family

Application07/251,149Parent of this continuation-in-part · filed 29 Sep 1988
Application07/404,721Parent of this continuation · filed 8 Sep 1989
Application07/876,804Parent of this division · filed 29 Apr 1992
US 2002/0192718 A1Pre-grant publication · published 19 Dec 2002
US 6,235,488 B1Parent of this division
Application07/876,804Parent of this continuation
Application07/404,721Parent of this continuation-in-part
US 5,130,257 AParent of this continuation-in-part
US 6,586,232 B2This patent

From the US filing history. Foreign family members aren’t listed.