Cleavage Coupled Lateral Flow Immunoassay for Rapid Endotoxin Testing

DESCRIPTION provided by applicant Bacterial endotoxins namely lipopolysaccharide LPS are common contaminants of many pharmaceutical medical device and food products that can elicit very strong immune responses in people and animals sometimes resulting in septic shock As such monitoring for endotoxin in medical and food products is essential and a $ billion annual market Current methods for detecting endotoxin are complex expensive and difficult to standardize Hence there is an unmet need for rapid simple assays that can be used to quickly assess endotoxin levels in field or point of care settings Lateral flow based assays LFAs are ideally suited for end users that require frequent rapid and straightforward endotoxin testing LFAs require very little sample no equipment or additional reagents and are simple robust and produced for $ each at scale We will develop a novel cleavage coupled competitive lateral flow assay C LFA that will enable inexpensive lateral flow devices that require no external instrumentation or skilled personnel to use The basis of the assay technology is a novel peptide construct that incorporates the cleavage site for an endotoxin activated protease between mimetope peptides that are specifically recognized by a particular monoclonal antibody The intact dimer form of this construct has high affinity for the antibody but the monovalent affinity of the cleaved form is considerably weaker Thus activity can unmask the antibodyandapos s antigen binding sites previously blocked by the dimer peptide construct by cleavage to low affinity monomers The specific aims of this Phase I application are to Optimize the dimer peptide and Develop the LPS C LFA prototype The long term goal of this project is to develop lateral flow immunoassays that can rapidly determine endotoxin levels above or below set thresholds We have previously developed these mimetope peptides as lateral flow based sensors for their cognate antibody and are thus well poised to leverage that technique for the unique problem of enzyme activity via lateral flow PUBLIC HEALTH RELEVANCE Bacterial endotoxins are common contaminants of many drugs medical devices and food products that can trigger very strong immune responses in people and animals sometimes resulting in septic shock We will develop a rapid simple and low cost lateral flow test that anyone can use to quickly monitor endotoxin levels in field or point of care settings with no need for equipment or additional reagents