Developing ARG1 and ODC Standardsfor a Microfluidic ELISA Device

Enzyme-Linked Immunosorbent 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 significant challenge. While microfluidic ELISAs requiring 2 μL 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 samples 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 overproducing 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.