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US 12,379,387 B2Granted
Engineered particles as red blood cell mimics and compositions containing same for hematology
AssigneeSlingshot Biosciences, Inc.
InventorsDaixuan Zhang, Jeffrey Kim, Keunho Ahn
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Abstract
Provided herein are red blood cell control compositions containing one or more populations of lysable hydrogel particles having an impedance that is substantially similar to the impedance of a human red blood cell of average diameter; and a population of hemoglobin molecules or a population of dye molecules that have substantially similar absorbance as hemoglobin; wherein (i) and/or (ii) are present in an amount that corresponds to a normal blood sample or disease state, or (i) and (ii) are present at a ratio that corresponds to a normal blood sample or disease state.
Claims23
A synthetic whole blood sample control, comprising:
- (i) a population of lysable hydrogel particles having an impedance that is substantially similar to the impedance of a human red blood cell of average diameter;
- (ii) a population of hemoglobin molecules and/or a population of dye molecules that has substantially similar absorbance as hemoglobin;
- and
- (iii) a population of non-lysable hydrogel particles having an impedance that is substantially similar to the impedance of a human white blood cell of average diameter,
- wherein the population of lysable hydrogel particles comprises a polymerized monomer and a co-monomer, wherein the co-monomer is a bifunctional monomer, and
- wherein the bifunctional monomer comprises a disulfide bond, an acid labile bond, or a combination thereof.
3 more elements
A method of calibrating a hematology analyzer for analysis of a target whole blood sample comprising red blood cells and white blood cells, the method comprising:
- (a) inserting into the hematology analyzer a synthetic whole blood sample control, wherein the synthetic whole blood sample control comprises:
- (i) a population of lysable hydrogel particles having an impedance that is substantially similar to the impedance of a human red blood cell of average diameter;
- (ii) a population of hemoglobin molecules and/or a population of dye molecules that has substantially similar absorbance as hemoglobin;
- and
- (iii) a population of non-lysable hydrogel particles having an impedance that is substantially similar to the impedance of a human white blood cell of average diameter,
- (b) measuring the impedance of the population of lysable hydrogel particles using the hematology analyzer;
- (c) calibrating the hematology analyzer for analysis of the red blood cells based on the measured impedance of the lysable hydrogel particles,
- (d) bringing the synthetic whole blood sample control in contact with a lysis buffer, wherein the population of lysable hydrogel particles comprise a bifunctional monomer comprising a bond that is capable of being lysed by the lysis buffer, thereby lysing the lysable population of hydrogel particles, and
- measuring the impedance of the population of non-lysable hydrogel particles using the hematology analyzer, and
- (e) calibrating the hematology analyzer for analysis of white blood cells in the target whole blood sample based on the measured impedance of the non-lysable hydrogel particles, and wherein:
- (i) the bifunctional monomer comprises a disulfide bond and wherein the lysis buffer is selected from the group consisting of dithiothreitol (DTT), tris(2-carboxyethyl)phosphine (TCEP), beta-mercaptoethanol (BME), glutathione (GSH), and combinations thereof,
- and/or
- (ii) the bifunctional monomer comprises an acid labile bond and wherein the lysis buffer comprises formic acid.
10 more elements
A method of evaluating a target whole blood sample comprising red blood cells and white blood cells, the method comprising:
- (a) inserting into a hematology analyzer a synthetic whole blood control, wherein the synthetic whole blood control comprises:
- (i) a population of lysable hydrogel particles having an impedance that is substantially similar to the impedance of a human red blood cell of average diameter;
- (ii) a population of hemoglobin molecules and/or a population of dye molecules that has substantially similar absorbance as hemoglobin;
- and
- (iii) a population of non-lysable hydrogel particles having an impedance that is substantially similar to the impedance of a human white blood cell of average diameter,
- and measuring the impedance of the population of the lysable hydrogel particles and the impedance of the population of the non-lysable hydrogel particles using the hematology analyzer,
- (b) inserting into the hematology analyzer the target whole blood sample and measuring the impedance of the red blood cells and the white blood cells in the target whole blood sample using the hematology analyzer,
- (c) comparing the impedance of the red blood cells in the target whole blood sample with the impedance of the population of the lysable hydrogel particles in the synthetic whole blood control, and comparing the impedance of the white blood cells in the target whole blood sample with the impedance of the population of the non-lysable hydrogel particles in the synthetic whole blood control,
- wherein the population of lysable hydrogel particles comprises a polymerized monomer and a co-monomer, wherein the co-monomer is a bifunctional monomer, and
- wherein the bifunctional monomer comprises a disulfide bond, an acid labile bond, or a combination thereof.
7 more elements
A method of evaluating a target whole blood sample comprising red blood cells and white blood cells, the method comprising:
- (a) inserting into a hematology analyzer a synthetic whole blood control, wherein the synthetic whole blood control comprises:
- (i) a population of lysable hydrogel particles having an impedance that is substantially similar to the impedance of a human red blood cell of average diameter;
- (ii) a population of hemoglobin molecules and/or a population of dye molecules that has substantially similar absorbance as hemoglobin;
- and
- (iii) a population of non-lysable hydrogel particles having an impedance that is substantially similar to the impedance of a human white blood cell of average diameter,
- and measuring the impedance of the population of lysable hydrogel particles using the hematology analyzer,
- (b) inserting into the hematology analyzer the target whole blood sample and measuring the impedance of the red blood cells in the target whole blood sample using the hematology analyzer,
- (c) comparing the impedance of the red blood cells in the target whole blood sample with the impedance of the population of the lysable hydrogel particles in the synthetic whole blood control,
- (d) bringing the synthetic whole blood control in contact with a lysis buffer, wherein the population of the lysable hydrogel particles comprise a bifunctional monomer comprising a bond that is capable of being lysed by the lysis buffer, thereby lysing the lysable population of hydrogel particles,
- (e) measuring the impedance of the population of the non-lysable hydrogel particles using the hematology analyzer,
- (f) measuring the impedance of the white blood cells in the target whole blood sample using the hematology analyzer, and
- (g) comparing the impedance of the white blood cells in the target whole blood sample with the impedance of the population of non-lysable hydrogel particles in the synthetic whole blood control,
- wherein:
- i) the bifunctional monomer comprises a disulfide bond and wherein the lysis buffer is selected from the group consisting of dithiothreitol (DTT), tris(2-carboxyethyl)phosphine (TCEP), beta-mercaptoethanol (BME), glutathione (GSH), and combinations thereof,
- or
- (ii) the bifunctional monomer comprises an acid labile bond and wherein the lysis buffer comprises formic acid.
13 more elements
Dependencies are inferred from the claim text.