Abstract
The country is experiencing a dramatic increase in the incidence of Diabetes Type II. In an effort to develop a medical device to both screen and monitor for Diabetes, we commenced a research project that embraced a team approach.
The overall objective was the design and development of a simple device that measures glycated hemoglobin, referred to as hemoglobin A1c, by electrochemical means. Patient blood can be accessed by finger prick, and after applying a drop of the blood to a device, lysis occurs by hemolyzing agents. Enzymatic activity will then perform the following steps: (a) digestion of the hemoglobin by a proteolytic enzyme, such that the glycated amino acid is liberated from the intact hemoglobin. (b) a second enzyme generates peroxide by oxidation of the glycated amino acid, (c) electrochemical detection of the H2O2.
Natalie Buzaglo investigated the lysis of whole blood cells using various hemolytic agents, which is needed to expose the glycated hemoglobin.
Ifaz Parvez prepared glycated hemoglobin by the non-enzymatic reaction of glucose with both crystalline human hemoglobin and horse hemoglobin. The glycated hemoglobin was purified by ion exchange chromatography. The presence of glycated hemoglobin was confirmed by MALDI TOF massspectroscopy.
Jose Cabrera investigated the enzyme kinetics for the proteinase step using Ifaz’s glycated product and evaluated the compatibility of the hemolytic agents by Natalie. Mass-spectroscopy, specifically, LC-MS confirmed proteinase activity by observation of the final substrate, fructosyl valine.
Oliver Moreira synthesized fructosyl valine, which can be used as a control for the enzymatic reactions.
Faculty Sponsors
Dr. Terry McCaffrey
Project Type
Event
Location
Alvin Sherman Library
Start Date
4-12-2013 1:00 PM
End Date
4-12-2013 5:30 PM
Model System for Design of a Medical Diagnostic Device: Hemoglobin A1c
Alvin Sherman Library
The country is experiencing a dramatic increase in the incidence of Diabetes Type II. In an effort to develop a medical device to both screen and monitor for Diabetes, we commenced a research project that embraced a team approach.
The overall objective was the design and development of a simple device that measures glycated hemoglobin, referred to as hemoglobin A1c, by electrochemical means. Patient blood can be accessed by finger prick, and after applying a drop of the blood to a device, lysis occurs by hemolyzing agents. Enzymatic activity will then perform the following steps: (a) digestion of the hemoglobin by a proteolytic enzyme, such that the glycated amino acid is liberated from the intact hemoglobin. (b) a second enzyme generates peroxide by oxidation of the glycated amino acid, (c) electrochemical detection of the H2O2.
Natalie Buzaglo investigated the lysis of whole blood cells using various hemolytic agents, which is needed to expose the glycated hemoglobin.
Ifaz Parvez prepared glycated hemoglobin by the non-enzymatic reaction of glucose with both crystalline human hemoglobin and horse hemoglobin. The glycated hemoglobin was purified by ion exchange chromatography. The presence of glycated hemoglobin was confirmed by MALDI TOF massspectroscopy.
Jose Cabrera investigated the enzyme kinetics for the proteinase step using Ifaz’s glycated product and evaluated the compatibility of the hemolytic agents by Natalie. Mass-spectroscopy, specifically, LC-MS confirmed proteinase activity by observation of the final substrate, fructosyl valine.
Oliver Moreira synthesized fructosyl valine, which can be used as a control for the enzymatic reactions.
