Towards Development Of Low Cost Electrochemical Biosensor For Detecting Percentage Glycated Hemoglobin
Siva Rama Krishna, V
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There is an ever growing demand for low cost biosensors in medical diagnostics. A well known commercially successful example is glucose biosensors which are used to diagonize and monitor diabetes. These biosensors use electrochemical analysis (electro analysis) as transduction mechanism. Electro analytical techniques involve application of electrical stimulus to the chemical/biochemical system under consideration and measurement of electrical response due to the oxidation and reduction reactions that occur because of the stimulus. They offer a lot of advantages in terms of sensitivity, selectivity, cost effectiveness and compatibility towards integration with electronics. Besides glucose, there are several biomolecules of significance for which electro analysis can potentially be used to develop low cost, rapid, easy to use biosensors. One such biomolecule is Glycated Hemoglobin (GHb). It is a post translational, non-enzymatic modification of hemoglobin with glucose and is a very good biomarker that indicates the average value of blood glucose over the past 120 days. It is always expresses as a percentage of total hemoglobin present in blood. Monitoring diabetes based on the value of percentage Glycated hemoglobin is advantageous as it gives an average value of glucose unlike plasma glucose values which vary a lot on a day to day basis depending on the dietary habits and the stress levels of the individual. This thesis is focused on the development of a low coat, easy to use, disposable sensor for measuring percentage Glycated hemoglobin. The first challenge in developing such a sensor is isolation of hemoglobin. Unlike glucose which is present in blood plasma (liquid content of blood), hemoglobin resides inside red blood cells also known as erythrocytes. O isolate hemoglobin, these cells have to be broken or lysed. All the existing approaches rely on mixing blood with lysing reagents to lyse erythrocytes. Ideal biosensors should be devoid of liquid reagents. Keeping this in perspective, in this thesis, this challenge is addressed by developing two entirely buffer/reagentless techniques to lyse erythrocytes and isolate hemoglobin. In the first technique, cellulose acetate membranes are embedded with lysing reagents and are used for lysing reagents and are used for lysing application. In the second techniques, commercially available nylon mesh nets are modified with lysing reagents to lyse and isolate hemoglobin. These membranes or mesh nets can be easily integrated on top of a disposable strip. After isolating hemoglobin, the next challenge is to selectively detect Glycated hemoglobin. Boronic acid conjugates are known to bind Glycated hemoglobin. Using this principle, a new composite is sysnthesized to specifically detect glc\ycated hemoglobin. The composite (GO-APBA) is a result of functionalization of Graphene Oxide (GO) with 3-aminophenylboronic acide (APBA). Detection of Glycated hemoglobin is achieved by modifying screen printed electrode strips with the synthesized compound, thus taking a step forwards achieving the objective. Since Glycated hemoglobin is always expressed as a percentage of hemoglobin, the next challenge is to detect total hemoglobin. In this thesis a low cost way of detecting hemoglobin is achieved by using GO modified or surfactant modified screen printed electrode strips. Furthermore, the potential interferences that blood plasma can cause in these measurements are eliminated with the help of permselective coatings. Thus using the technologies developed in this thesis, measurements of percentage Glycated hemoglobin can be potentially made on handheld electronic devices akin to glucose meters by using just a drop of blood.