Quantitation of functional groups on solid supports
AssigneeSiemens Healthcare Diagnostics Inc.
InventorsSpencer Lin, Christopher Reamer, Ryan Spears
Life of this patent
Estimating the term…
Abstract
Processes for quantifying an amount of functional groups immobilized on a solid support are described herein. The processes allow for determining whether sufficient functional groups are provided on a solid support for the attachment of a first binding pair member for the detection of a target analyte.
Claims20
A method for quantifying target functional groups immobilized on a solid support comprising one or more paramagnetic particles, the method comprising the steps of:
- (a) contacting the target functional groups immobilized on the one or more paramagnetic particles with an amount of a selective compound, wherein an amount of the selective compound binds directly to the target functional groups and forms bound selective compound and an amount of the selective compound does not bind to the target functional groups and is present as unbound selective compound, wherein the target functional groups comprise aldehydic functional groups, and wherein the selective compound comprises propylamine;
- (b) separating a supernatant from the paramagnetic particles, wherein the supernatant contains an amount of the unbound selective compound;
- (c) adding an indicator to the supernatant which binds to unbound selective compound in the supernatant to provide a colorimetric result, wherein the indicator comprises a solution comprising 2,4,6-trinitrobenzene sulfonic acid (TNBSA);
- (d) measuring the colorimetric result;
- (e) determining from the measured result an amount of unbound selective compound in the supernatant;
- (f) determining an amount of bound selective compound immobilized on the one or more paramagnetic particles from the amount of unbound selective compound;
- and
- (g) determining from the amount of the immobilized selective compound an amount of the target functional groups immobilized on the one or more paramagnetic particles.
A method for quantifying target functional groups immobilized on a solid support comprising one or more paramagnetic particles, the method comprising:
- (a) contacting the target functional groups immobilized on the one or more paramagnetic particles with an amount of a selective compound, wherein an amount of the selective compound binds directly to the target functional groups and forms bound selective compound and an amount of the selective compound does not bind to the target functional groups and is present as unbound selective compound, wherein the target functional groups comprise amino functional groups, and wherein the selective compound comprises an aldehydic compound which selectively reacts with immobilized amino functional groups on the one or more paramagnetic particles;
- (b) separating a supernatant from the paramagnetic particles, wherein the supernatant contains an amount of the unbound selective compound;
- (c) adding an indicator to the supernatant which binds to unbound selective compound in the supernatant to provide a colorimetric result, wherein the indicator comprises 4-Amino-3-hydrazino-5-mercapto-1,2,4-triazole;
- (d) measuring the colorimetric result;
- (e) determining from the measured result an amount of unbound selective compound in the supernatant;
- (f) determining an amount of bound selective compound immobilized on the one or more paramagnetic particles from the amount of unbound selective compound;
- and
- (g) determining from the amount of the immobilized selective compound an amount of the target functional groups immobilized on the one or more paramagnetic particles.
Dependencies are inferred from the claim text.