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US 12,140,596 B2Granted
Kits, microfluidics devices, and methods for performing biotin assays
AssigneeSiemens Healthcare Diagnostics Inc.
InventorsTie Q Wei, Zhu Teng, Xiaodong Liu, Martin Drinan
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Abstract
Kits containing a chemiluminescent detection system and microfluidics devices and methods for determining the concentration of biotin in a sample are disclosed. The kits, microfluidics devices, and methods utilize a sensitizer capable of generating singlet oxygen in its excited state and having avidin or an analog thereof directly or indirectly bound thereto, as well as a singlet oxygen-activatable chemiluminescent compound having biotin or an analog thereof directly or indirectly bound thereto.
Claims13
A kit containing a chemiluminescent detection system for determining the concentration of biotin in a sample, the kit comprising:
- (a) a composition comprising a particle having a singlet oxygen-activatable chemiluminescent compound coupled thereto and/or disposed therein, the particle having biotin or an analog thereof directly or indirectly bound thereto, wherein the singlet oxygen-activatable chemiluminescent compound is a substance that undergoes a chemical reaction with singlet oxygen to form a metastabile intermediate species that can decompose with the simultaneous or subsequent emission of light;
- and
- (b) a composition comprising a particle having a sensitizer capable of generating singlet oxygen in its excited state coupled thereto and/or disposed therein, the particle having traptavidin directly or indirectly bound thereto;
- and
- wherein the kit is capable of detecting biotin concentrations in a range of from about 0 to about 1500 ng/ml;
- wherein the biotin analog is selected from the group consisting of 4′-hydroxyazobenzene-2-carboxylic acid (HABA), iminobiotin, biotin carbonate, biotin carbamate, biotin methyl ester, desthiobiotin, diaminobiotin, chloroacetylated biotin, biotin sulfone, thiobiotin, methoxycarbonylbiotin methyl ester, bis-biotin, tetra-biotin, and an ester or salt of any of the above biotin analogs.
3 more elements
A kit containing a chemiluminescent detection system for determining the concentration of biotin in a sample, the kit comprising:
- (a) a composition comprising a particle having a singlet oxygen-activatable chemiluminescent compound coupled thereto and/or disposed therein, the particle having 4′-hydroxyazobenzene-2-carboxylic acid (HABA) directly or indirectly bound thereto;
- and
- (b) a composition comprising a particle having a sensitizer capable of generating singlet oxygen in its excited state coupled thereto and/or disposed therein, the particle having traptavidin directly or indirectly bound thereto;
- wherein the kit is capable of detecting biotin concentrations in a range of from about 0 to about 1500 ng/ml;
- wherein the singlet oxygen-activatable chemiluminescent compound is a substance that undergoes a chemical reaction with singlet oxygen to form a metastabile intermediate species that can decompose with the simultaneous or subsequent emission of light.
2 more elements
A method for detecting the presence and/or concentration of biotin in a sample, the method comprising the steps of:
- (a) combining, either simultaneously or wholly or partially sequentially:
- (1) a sample suspected of containing biotin;
- (2) a composition comprising a particle having a singlet oxygen-activatable chemiluminescent compound coupled thereto and/or disposed therein, the particle having biotin or a biotin analog directly or indirectly bound thereto, wherein the singlet oxygen-activatable chemiluminescent compound is a substance that undergoes a chemical reaction with singlet oxygen to form a metastabile intermediate species that can decompose with the simultaneous or subsequent emission of light;
- and
- (3) an excess of a composition comprising a particle having a sensitizer capable of generating singlet oxygen in its excited state coupled thereto and/or disposed therein, the particle having traptavidin directly or indirectly bound thereto;
- (b) allowing the binding of (3) to biotin present in the sample or to (2), whereby in the absence of biotin, a complex is formed between (2) and (3) and the sensitizer is brought into close proximity to the chemiluminescent compound;
- (c) activating the sensitizer to generate singlet oxygen, wherein activation of the sensitizer present in the complex causes the activation of the chemiluminescent compound present in the complex;
- (d) determining the amount of chemiluminescence generated by the activated chemiluminescent compound;
- (e) optionally repeating steps (b)-(d);
- and
- (f) detecting the presence and/or concentration of biotin by analyzing the amount of chemiluminescence so produced, wherein the amount of chemiluminescence is inversely proportional to the amount of biotin in the sample;
- and
- wherein the method is capable of detecting biotin concentrations in a range of from about 0 to about 1500 ng/ml;
- wherein the biotin analog is selected from the group consisting of 4′-hydroxyazobenzene-2-carboxylic acid (HABA), iminobiotin, biotin carbonate, biotin carbamate, biotin methyl ester, desthiobiotin, diaminobiotin, chloroacetylated biotin, biotin sulfone, thiobiotin, methoxycarbonylbiotin methyl ester, bis-biotin, tetra-biotin, and an ester or salt of any of the above biotin analogs.
11 more elements
Dependencies are inferred from the claim text.