| Summary: | 碩士 === 國立臺灣大學 === 醫學工程學研究所 === 97 === Immunoassays are bioanalytical methods which detect and quantify the concentration of a certain analyte through the specific affinity between an antigen (analyte) and an antibody. Immunoassays have been widely used in many important areas of pharmaceutical analysis, such as diagnosis of diseases, therapeutic drug monitoring, clinical pharmaco¬kinetic, etc. Immunoassays are achieved by measuring the label activity (e.g. radiation, fluorescence, or enzyme) of antigens or antibodies. However, immunoassays often require special apparatus and instruments, which are expensive and time-consuming. Furthermore, signal detection of immunoassays is also easily disturbed by non-specific bindings.
Therefore, the purpose of this study focuses on reducing experimental expenses and non-specific bindings of immunoassays. Utilizing its porous structure and the easily modifiable surface, bamboo charcoal can be modified to become a suitable material for immunoassays. In addition, we designed a new immunoassay device in order to reduce the complexity and expenses of the experiment.
There are many advantages with using bamboo charcoal and polyoxyethylene bis-amine (PEGBA) as the main experimental materials. First, the porous surface of bamboo charcoal gives rise to a large surface area. Also, the abundant carbon double bonds on surface of bamboo charcoal are easy to be modified. By treating bamboo charcoal with acid, the surface will become full of carboxyl groups, which then can bind with the amino groups of PEGBA through amide bonds. While one end of PEGBA is grafted on the surface of bamboo charcoal, the other end is able to bind with the C-terminus of antibodies, leaving the N-terminus available to antigens. Moreover, the special structure of PEGBA can lead to a waving effect, which can cause steric repulsion and reduce non-specific bindings. This method can improve the accuracy of immunoassays.
In conclusion, bamboo charcoal can be modified through carboxylation and PEGBA grafting. Modified bamboo charcoal can then be successfully conjugated with antibodies and become a multiplexed immunoassay material. Comparing with commercial magnetic nanoparticles, our device is more effective in reducing non-specific bindings and saving experimental expenses and time.
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