University of Louisville
Lyoprotectants for Stabilizing Hemoglobin and Extending Red Blood Cell Shelf Life
Grade Level at Time of Presentation
Junior
Major
Biology
2nd Grade Level at Time of Presentation
Junior
2nd Student Major
Biology
Institution 23-24
University of Louisville
KY House District #
42
KY Senate District #
35
Faculty Advisor/ Mentor
Dr. Michael Menze, PhD; Mr. Charles Elder
Department
Dept. of Biology
Abstract
Extending the shelf-life of red blood cells (RBCs) and allowing for ambient storage through dry preservation would significantly reduce casualties during blood shortages worldwide. Currently, RBCs last a mere 35-42 days at 4 °C post-donation. Two primary forms of damage to RBCs during drying are RBC hemolysis and hemoglobin oxidation. Hemolysis is the leakage of hemoglobin into the surrounding solution caused by RBC lysis or pore formation in the cell membrane. Hemoglobin oxidation occurs when the iron within the heme group of hemoglobin oxidizes from Fe2+ to Fe3+, forming methemoglobin.Methemoglobin cannot bind oxygen, reducing the therapeutic value of RBC transfusion units. My research focuses on preventing hemoglobin oxidation during drying to maintain RBC functionality. I am studying compounds produced in organisms that can survive complete drying, such as sugars, proteins, and polymers, to inhibit hemoglobin oxidation during drying. Our project involves lysing porcine RBCs, isolating the oxygen transport protein hemoglobin, and preparing samples with different protectant-to-hemoglobin ratios. After freeze-drying and rehydrating the hemoglobin, oxidation levels are quantified spectrophotometrically. Utilizing this method, we can determine the concentration of the protectant in which fifty percent less of the hemoglobin becomes oxidized. These values are significant between sugars when analyzed based on total molecules present during drying but do not show significance when analyzed based on total weight. With further investigation,we hope to identify the material and chemical properties of these protectants, which allow them to inhibit hemoglobin oxidation during drying. Elucidating the protective mechanisms will enable the development of dry preservation techniques that maximize RBC shelf-life for enhanced availability and transportation.
Lyoprotectants for Stabilizing Hemoglobin and Extending Red Blood Cell Shelf Life
Extending the shelf-life of red blood cells (RBCs) and allowing for ambient storage through dry preservation would significantly reduce casualties during blood shortages worldwide. Currently, RBCs last a mere 35-42 days at 4 °C post-donation. Two primary forms of damage to RBCs during drying are RBC hemolysis and hemoglobin oxidation. Hemolysis is the leakage of hemoglobin into the surrounding solution caused by RBC lysis or pore formation in the cell membrane. Hemoglobin oxidation occurs when the iron within the heme group of hemoglobin oxidizes from Fe2+ to Fe3+, forming methemoglobin.Methemoglobin cannot bind oxygen, reducing the therapeutic value of RBC transfusion units. My research focuses on preventing hemoglobin oxidation during drying to maintain RBC functionality. I am studying compounds produced in organisms that can survive complete drying, such as sugars, proteins, and polymers, to inhibit hemoglobin oxidation during drying. Our project involves lysing porcine RBCs, isolating the oxygen transport protein hemoglobin, and preparing samples with different protectant-to-hemoglobin ratios. After freeze-drying and rehydrating the hemoglobin, oxidation levels are quantified spectrophotometrically. Utilizing this method, we can determine the concentration of the protectant in which fifty percent less of the hemoglobin becomes oxidized. These values are significant between sugars when analyzed based on total molecules present during drying but do not show significance when analyzed based on total weight. With further investigation,we hope to identify the material and chemical properties of these protectants, which allow them to inhibit hemoglobin oxidation during drying. Elucidating the protective mechanisms will enable the development of dry preservation techniques that maximize RBC shelf-life for enhanced availability and transportation.