Methods and compositions for quantifying hemoglobin
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Publication Number
US-11353466-B2
Publication Date
2022-06-07
Expiration Date
Abstract
The present invention relates to methods and compositions for quantifying hemoglobin using, inter alia, a peroxidase substrate and hydrogen peroxide.
Core Innovation
The document describes a method for quantifying hemoglobin in a sample by contacting the sample with a peroxidase substrate and hydrogen peroxide, under suitable conditions, to form an oxidation product. The hemoglobin level is determined by assessing the peroxidase substrate and/or the oxidation product. The peroxidase substrate is oxidized by hydrogen peroxide to generate an oxidation product, including an example oxidation product identified as 7-hydroxyphenoxazin-3-one.
The sample analyzed in the method comprises a cellular membrane coated nanoparticle, where the nanoparticle includes an inner core comprising a non-cellular material and an outer surface comprising a cellular membrane derived from a hemoglobin-containing cell. The document describes this nanoparticle architecture in connection with hemoglobin-containing cellular membrane-coated nanoparticles, and the sample may further include a releasable cargo.
The disclosed peroxidase substrate and detection approach includes substrates and readouts such as ADHP and detection by colorimetric and/or fluorescent assessment of the oxidation product. The document further describes a kit/system configuration for quantifying hemoglobin that comprises the peroxidase substrate and hydrogen peroxide, and a solution or suspension having basic pH and/or containing a surfactant for preparing the sample.
Claims Coverage
The document includes two independent claims, one directed to a hemoglobin quantification method and one directed to a hemoglobin quantification kit. Across these independent claims, the inventive subject matter centers on a peroxidase substrate/hydrogen peroxide oxidation assay coupled to assessment of the oxidation product, and on using a specific sample type: a cellular membrane coated nanoparticle with a non-cellular inner core and a cellular membrane derived from a hemoglobin-containing cell. An additional independent-claim distinction is that the kit claim includes a solution or suspension with basic pH and/or surfactant as part of the kit components.
Peroxidase substrate oxidation and assessment for hemoglobin quantification
Contacting a sample containing or suspected of containing hemoglobin with a peroxidase substrate and hydrogen peroxide to form an oxidation product, and assessing the peroxidase substrate and/or the oxidation product to quantify hemoglobin.
Cellular membrane coated nanoparticle sample architecture
Using a sample that comprises a cellular membrane coated nanoparticle having an inner core comprising a non-cellular material and an outer surface comprising a cellular membrane derived from a hemoglobin-containing cell.
Kit components for hemoglobin quantification
Providing a kit comprising a peroxidase substrate and hydrogen peroxide, together with a solution or suspension having basic pH and/or containing a surfactant for preparing a sample.
Kit usage with cellular membrane coated nanoparticle
Wherein the sample in the kit context comprises a cellular membrane coated nanoparticle having an inner core comprising a non-cellular material and an outer surface comprising a cellular membrane derived from a hemoglobin-containing cell.
Overall claim coverage focuses on hemoglobin quantification via peroxidase-mediated oxidation (peroxidase substrate plus hydrogen peroxide with assessment of the substrate and/or oxidation product), and on performing the quantification on a cellular membrane coated nanoparticle sample defined by a non-cellular inner core and an outer cellular membrane derived from a hemoglobin-containing cell. The kit claim additionally requires inclusion of basic pH and/or surfactant-containing solution or suspension as part of the kit components.
Stated Advantages
Not explicitly described in patent.
Documented Applications
Not explicitly described in patent.
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