Determining the density of functional moieties on polymer reagents
First Claim
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1. A method of determining the density of functional moieties on polymer substrates comprising:
- (a) attaching a dye molecule to a polymer substrate, said dye molecule absorbing infrared radiation at a wavelength of at least 600 nm;
(b) attaching a compound as a functional moiety to said polymer substrate with attached dye molecules, said compound being selected to react with an analyte in a biological sample;
(c) attaching the dye molecule selected in (a) as a label to a second sample of said polymer substrate and determining by their absorption of infrared radiation the number of dye molecules attached to said polymer substrate;
(d) determining the absorption of ultraviolet radiation by the functional moieties and by infrared radiation absorption of dye molecules in the polymer substrate of (b); and
(e) determining the number of functional moieties on said polymer substrate of (b) relative to the number of attached dye molecules determined in (c), said number of functional moieties representing the density of said functional molecules.
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Abstract
The invention provides a method for determining the density of functional molecules attached to a substrate used for analysis of biological samples. A dye molecule responding to near infrared radiation at a wavelength of at least 600 nm is attached to the substrate and used to indicate the number of the functional molecules attached to the substrate by comparing the infrared absorption of the dye molecules with the ultraviolet absorption of the functional molecules. Such substrates may be employed in immunoassays and in vivo diagnostics.
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Citations
15 Claims
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1. A method of determining the density of functional moieties on polymer substrates comprising:
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(a) attaching a dye molecule to a polymer substrate, said dye molecule absorbing infrared radiation at a wavelength of at least 600 nm;
(b) attaching a compound as a functional moiety to said polymer substrate with attached dye molecules, said compound being selected to react with an analyte in a biological sample;
(c) attaching the dye molecule selected in (a) as a label to a second sample of said polymer substrate and determining by their absorption of infrared radiation the number of dye molecules attached to said polymer substrate;
(d) determining the absorption of ultraviolet radiation by the functional moieties and by infrared radiation absorption of dye molecules in the polymer substrate of (b); and
(e) determining the number of functional moieties on said polymer substrate of (b) relative to the number of attached dye molecules determined in (c), said number of functional moieties representing the density of said functional molecules. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9)
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- 10. A method of carrying out immunoassays wherein a polymer substrate attached to a functional molecule and a dye molecule having the density of said functional molecules determined by the number of attached dye molecules is used as a reagent for determining analytes of interest in biological samples.
- 13. A method of carrying out in vivo diagnostics wherein a polymer substrate is used as a reagent for imaging of predetermined species of body cells, said polymer substrate being attached to a functional molecule and a dye molecule and the density of said functional molecules is determined by the number of attached dye molecules.
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15. An immunoassay wherein a functional moiety is attached to a polymer substrate and thereafter contacted with a biological sample for binding said functional moiety to an analyte, comprising:
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(a) attaching a dye molecule to a polymer substrate, said dye molecule absorbing infrared radiation at a wavelength of at least 600 nm;
(b) attaching a functional moiety to said polymer substrate having an attached dye molecule of (a), said functional moiety being selected to react with an analyte in a biological sample;
(c) attaching the dye molecules of (a) as a label to a second sample of said polymer substrate and determining by their absorption of radiation the number of dye molecules attached to said polymer substrate;
(d) determining the absorption of radiation by the functional moieties and dye molecules in the polymer substrate of (b);
(e) determining the number of functional moieties in said polymer substrate of (b) relative to the number of attached dye molecules determined in (c). (f) contacting with said biological sample the polymer substrate of (b) having said functional moiety and said dye molecule attached under suitable conditions to react said functional moiety with said analyte;
(g) measuring the absorption by radiation of the combined polymer substrate and biological sample of (f); and
(h) determining the amount of the analyte which is bound to the functional moiety by the amount of said dye molecule present.
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Specification