Department

Chemistry

First Advisor

Dr. Debashis Dutta

Description

Enzyme - Linked Immunosorb e nt Assay , or ELISA , is a commonly used analytical technique that allows the quantitation of proteins in biological samples at very low concentrations. However, implementation of quantitative ELISAs using extremely small volumes of complex tissue samples still remains a signi ficant challenge. While microfluidic ELISAs requiring 2μL of the sample have recently been developed in Prof. Dutta’s laboratory, it’s been challenging to apply them to assess the levels of arginase 1 (ARG1) and ornithine decarboxylase (ODC) in tissue sam ples from mouse heart in the absence of any reliable standards. My project focused on preparing these standards to contain known amounts of the two enzymes which will be applied to quantitate our target microfluidic ELISAs. This is being accomplished by ov erproducing histidine - tagged ARG1 and ODC in E. coli and then purifying the enzymes using a nickel column. Future research will use these standards to estimate the amounts of ARG1 and ODC amounts in tissue samples from mouse heart.

Comments

Oral presentation, Wyoming NSF EPSCoR

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Developing ARG1 and ODC Standards for a Microfluidic ELISA Deviceice

Enzyme - Linked Immunosorb e nt Assay , or ELISA , is a commonly used analytical technique that allows the quantitation of proteins in biological samples at very low concentrations. However, implementation of quantitative ELISAs using extremely small volumes of complex tissue samples still remains a signi ficant challenge. While microfluidic ELISAs requiring 2μL of the sample have recently been developed in Prof. Dutta’s laboratory, it’s been challenging to apply them to assess the levels of arginase 1 (ARG1) and ornithine decarboxylase (ODC) in tissue sam ples from mouse heart in the absence of any reliable standards. My project focused on preparing these standards to contain known amounts of the two enzymes which will be applied to quantitate our target microfluidic ELISAs. This is being accomplished by ov erproducing histidine - tagged ARG1 and ODC in E. coli and then purifying the enzymes using a nickel column. Future research will use these standards to estimate the amounts of ARG1 and ODC amounts in tissue samples from mouse heart.