Immunoliposome Polymerase Chain Reaction (ILPCR) Assay

A simple-to-make diagnostic immunoassay capable of sub-femtomolar sensitivity that is very robust against matrix contaminations. Background and Technology: The US Army has developed an ultrasensitive immunoassay capable of sub-femtomolar (10-15) limit of detection. ILPCR technology overcomes the limitations of traditional immuno-PCR by encapsulating the signal amplification DNA/reporter inside a liposome, and incorporating a receptor (e.g. biotin or gangliosides) as the detection reagent into the bilayer of the liposome. After immobilization of the target analyte by the specific capture molecule and co-binding of the detection reagent, the liposomes are ruptured to release the reporters, which are quantified by real-time PCR. Preparation of the ILPCR reagent is simple. Unlike traditional immuno-PCR, synthesis of an antibody/DNA conjugate is not needed. By encapsulating the reporter inside the liposome, the reporter is protected by the liposome from degradation by impurities, which substantially reduces the possibility of false-negative results, and enables the use of DNase I to all but eliminate DNA contamination and false-positives. ILPCR’s improved sensitivity is due in part to the high number of reporters per binding event and the low nonspecific binding of the reagents. The assay has shown 5 orders of magnitude in dynamic range. Benefits:Ultrasensitive: Demonstrated sub-attomolar (10-18) and sub-femtomolar (10-15) detection threshold capabilities under ideal and “real-world” conditions respectivelyBroad Dynamic Range: Five Orders of Magnitude Accurate: Protection of the reporter inside the liposome greatly reduces false-negatives; eliminates DNA contamination, hence substantially reduces false-positives Versatile: Reduced to practice in biotoxins (botulinum and cholera), oncology biomakers (CEA), and viral HIV detection systems; applicable to a multitude of other analytes of commercial interest Simple: Reagents are easy to manufacture, inexpensive, and have a long shelf-life Status and Opportunity: Available for license and commercialization:One issued US patent (7,582,430), one published US patent (2009/0176250), and various pending foreign filings (WO2005/067583) Peer-reviewed journal articles available; additional information under non-disclosure agreement Potential for R&D; collaboration with inventors Dr. Tim O'Leary & Dr. Jeff Mason