Abstract
An immobilized liposome electrode (ILE)-based sensor was developed to quantify conformational changes of the proteins under various stress conditions. The ILE surface was characterized by using a tapping-mode atomic force microscopy (TM-AFM) to confirm surface immobilization of liposome. The uniform layer of liposome was formed on the electrode. The current deviations generated based on the status of the proteins under different stress were then measured. Bovine carbonic anhydrase (CAB) and lysozyme were tested with three different conditions: native, reduced and partially denatured. For both proteins, a linear dynamic range formed between denatured concentrations and output electric current signals was able to quantify conformational changes of the proteins. The pattern recognition (PARC) technique was integrated with ILE-based sensor to perform data analysis and provided an effective method to improve the prediction of protein structural changes. The ILE-based stress sensor showed potential of leveraging the amperometric technique to manifest activity of proteins based on various external conditions.
Highlights
Given that interest to fast, reliable and continuous measurements of biological molecules has been rising in medicine, biotechnology and environmental sciences, a biosensor which facilitates point-of-use with high sensitivity and affordability has been widely recognized as one of the promising alternatives in life science [1,2,3]
To achieve aforementioned prediction of protein structural changes, we describe the analytical characteristics of the resultant amperometric immobilized liposome electrode (ILE)-based sensor due to the electrolyte release from liposomes in the presence of proteins under various denaturant concentrations
After the self-assembled monolayer (SAM) were prepared on Au electrode, the liposomes were expected to be immobilized on the surface based on amino coupling
Summary
Given that interest to fast, reliable and continuous measurements of biological molecules has been rising in medicine, biotechnology and environmental sciences, a biosensor which facilitates point-of-use with high sensitivity and affordability has been widely recognized as one of the promising alternatives in life science [1,2,3]. There are several methods including array-type protein chip [20], florescence intensity [21] and circular dichroism (CD) [22] measurements to monitor stress-induced structural changes of proteins, those are expensive and labor-consuming. Those methods show the limitations for predicting the dynamics, structure deformation or interaction of proteins. The measurement of the markers can be used as a secondary response for monitoring protein-lipid membrane interactions, which are primary detection scheme used for the ILE-based sensor. We analyzed the response of ILE to proteins with pattern recognition (PARC) to improve the prediction of protein structural changes
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