A New reagent for detection of program cell death
Programmed cell death (PCD) like cell proliferation is an essential process of life. A novel reagent, a conjugate of anti- CT Receptor antibody and a fluorophore, has been studied and is rapidly internalised into live cells undergoing PCD (apoptosis, autophagy) in vitro. A similar reagent is being developed for detection of PCD in diseased tissues in vivo.
Our group has discovered that a novel reagent, a conjugate of anti-CTR antibody with a fluorophore AlexaFluor 568 is rapidly accumulated into cells cultured in vitro that have been induced to undergo autophagy by starvation or apoptosis with cytotoxin. This prototype has been succeeded conjugates made with pH dependent dyes such as pHrodo Red. The result is a high fluorescence signal that can be recorded with technologies such as confocal microscopy, flow cytometry and operetta platforms. This event overlaps the binding of annexin V and the activation of caspase 3, events previously used to determine PCD. As the antibody binds an extracellular epitope of CTR the rapid internalisation and concentration in the peri-nuclear region represents a novel event specific for PCD. The utility of similar radioactive conjugates has been proposed to image PCD in vivo and this might be useful for monitoring drug trials for the treatment of tumours and cardiovascular disease.
- Dr Peter Wookey
- Pragya Gupta – PhD student
Postdoc support from the Australian government, philanthropic funds, funds from the pharmaceutical industry and small institutional grants.
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