Amplified Detection of Enzymes without Biological Antibodies Sanku Mallik, North Dakota State University Fargo, DMR 0705767.
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Usually, small amounts of pathogenic enzymes are detected employing ELISA. Although this technique is very sensitive, it requires the use of biological antibodies. In addition to the expenses involved, since the antibodies are large biomolecules, proper handling and storage are required to maintain the activity. We are working on a new methodology for amplified detection of enzymes (similar to ELISA) without the use of biological antibodies. We have demonstrated the proof-of-concept by detecting low amounts (40 ng) of the cancer associated enzyme matrix metalloproteinase-9 (MMP-9) by using appropriately formulated liposomes.
The scheme for the amplified detection of MMP-9 without using biological antibodies. A rapid formation of brown color was observed in the presence of 40 ng of the enzyme; the assay solution remains clear in the absence of the enzyme.
Composition of the liposomes: Liposomes were prepared in 25 mM HEPES buffer, pH = 8.0 with total lipid concentration of 1 mg/mL. The structures of the lipids used in the liposomes are shown below.