PROXIMITY LIGATION ASSAY: A NEW TOOL FOR PROTEIN DETECTION
Kruthika P., Nagashree N. Rao and Ashwani Sharma*
ABSTRACT
The field of proteomics is growing at a very fast pace and new techniques are being introduced almost every year and modifications to the existing ones are underway. Proximity ligation Assay (PLA) is one such immunochemical method to detect proteins, protein-protein interactions and post translational modifications. The technique is based on the interaction of epitope of the target protein and antibodies. Antibodies bound to oligonucleotides that form proximity probes, which when in close proximity undergo a ligation event followed by amplification by PCR or Rolling circle amplification to produce a detectable signal. The assay is highly sensitive, with detection ranges in zeptamolar levels and specific as it is enabled by antigen-antibody interaction. With different variants, the technique will help in native and localized detection of protein-protein interactions as in the case of in-situ PLA and even protein-nucleotide interactions as in the case of ISH-PLA. Going forward this review also emphasizes on varied applications in multiple areas like cancer biology, neuroscience, diagnostics and pharmacology. Upcoming variants like multiplex PLA and 3PLA are also discussed giving insights about potential revolutions this assay could bring in the near future.
Keywords: PLA and 3PLA.
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