Assay and method for quantitating carbonic anhydrase activity and assessing red blood hemolysis
AssigneeCase Western University
InventorsRyan R. Geyer, Pan Zhao, Mark D. Parker, Walter F. Boron
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Abstract
A method and assay for using carbonic anhydrase (CA), particularly CA-I or CA-II, as a biomarker of hemolysis. The method and assay detect hemolysis by determining a percentage erythrocyte hemolysis in a specimen or sample of blood based upon quantification of carbonic anhydrase present in the extracellular portion of the blood. The method and test serve to optimize therapeutic efficacy for treatments of hemolysis.
Claims18
A method of measuring the amount or activity of carbonic anhydrase enzyme, in a bodily sample, the method comprising:
- mixing a bodily sample including RBC lysate with a first physiological CO2/HCO3− solution, the first physiological CO2/HCO3− solution having a pH of about 7;
- mixing the first physiological CO2/HCO3− solution, which includes the bodily sample, with a second CO2/HCO3− solution, which includes NaHCO3, having a basic dissimilar pH;
- and
- measuring the rate at which the pH of the newly mixed solution equilibrates under the influence of the enzyme to determine carbonic anhydrase activity.
A method for detecting red blood cell hemolysis in a bodily sample, the method comprising:
- mixing a bodily sample including red blood cells (RBC) and/or RBC lysate with a first physiological CO2/HCO3− solution, the first physiological CO2/HCO3− solution having a pH of about 7;
- mixing the first physiological CO2/HCO3− solution, which includes the bodily sample, with a second CO2/HCO3− solution, which includes NaHCO3, having a basic dissimilar pH;
- and
- measuring the rate at which the pH of the newly mixed solution equilibrates under the influence of the carbonic anhydrase enzyme in the bodily sample to determine red blood cell hemolysis in the bodily sample.
A method for determining red blood cell hemolysis in a bodily sample, the method comprising:
- mixing a bodily sample including red blood cells (RBC) and/or RBC lysate with a first physiological CO2/HCO3− solution, the first physiological CO2/HCO3− solution having a pH of about 7;
- mixing the first physiological CO2/HCO3− solution with a second CO2/HCO3− solution having a pH of about 8.4;
- measuring the rate at which the pH of the newly mixed solution equilibrates under the influence of the carbonic anhydrase enzyme in the bodily sample to determine red blood cell hemolysis in the bodily sample.
Dependencies are inferred from the claim text.