Olson Research Group:  Biochemistry

ELISA Development

New Sensitive and selective Enzyme-Linked Immunosorption Assays (ELISAs) are being developed in our laboratory. These will be used to analyze biomolecules present in small quantities in blood samples, urine samples, etc.

In a competitive ELISA, an antibody specific for a molecule of interest (antigen) is first affixed to a solid support, often the well of a microplate.
The well is next exposed to the sample. If present, the antigen will bind to the antibody. The plate is then washed thoroughly.
Next an antigen-enzyme conjugate is applied to the well. The conjugate will bind any free antibody sites on the plate. If the original sample contains few antigen molecules, many conjugates will bind (left series of wells). If the sample contains many antigen molecules, few conjugates can bind (right series of wells). The plates are again washed thoroughly.
Finally, a substrate for the enzyme is applied to the well. This substrate is chosen to produce a detectable product when acted upon by the enzyme. The quantity of the product can then be correlated to the concentration of the antigen in the sample.

• Our laboratory is investigating the use of different enzymes for ELISAs. Some optimal characteristics for an enzyme in this application are that it be economical, easy to purify, and have a high turnover number. Stability with regard to temperature, pH, and variable reaction conditions is also highly important. For example, enzymes that can function in non-aqueous environments have promise for ELISAs in organic solvents.

• We are also interested in designing new substrate molecules that, when converted to product, boost signal detection. This research incorporates a large amount of synthetic organic chemistry, and is being carried out in collaboration with Dr. Ippoliti’s laboratory at UST.

Please send me an email if you are interested in joining with my group in this work.