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Principal Investigator: Alfredo QUIJANO RUBIO
Organization: MONOD BIO, INC.
Fiscal Year: 2024
Award: $290,066
Funding agency: National Center for Advancing Translational Sciences
Abstract. Bacterial endotoxins are components of the outer cell membranes of gram-negative organisms and is
one of the most potent human toxins. Due to the health dangers associated with endotoxin contamination, testing
is critical for pharmaceuticals, biological products, and medical devices. The majority of endotoxin testing still
relies on the Limulus Amebocyte Lysate (LAL) assays using an extract from the blood of the horseshoe crabs.
This decades-old technology threatens a limited biological resource, which is considered vulnerable by
international organizations. We seek to enable the design of new endotoxin sensors with no requirement for
horseshoe crab blood or any of its components. In addition, unlike typical analytical assays composed of multiple
biological components and often complicated assay steps, our one-step lucCage biosensor can eliminate many
complexities of existing assays as it is a "single-step, all-in-solution" assay. Our approach will be enabled through
the (i) computational design and experimental selection of functional LPS bioluminescent biosensors; (ii) in vitro
evolution of biosensors to optimize sensitivity and stability; and (iii) optimization of the LPS detection assay
conditions and formulation of a 96-well kit. With our proprietary technology, we can develop alternative de novo
protein endotoxin biosensors that provide multiple advantages over current techniques: (i) sensitivity by using
bioluminescent readouts over turbidimetric, colorimetric or fluorescent readouts, (ii) broad dynamic range in a
single test that will reduce the total number of tests required by the user to quantify endotoxin in a sample, (iii)
stability and cost, as our biosensor proteins are highly stable and easy to manufacture. Thus, successful
development of endotoxin sensing lucCage biosensors will enable the development of sustainable, inexpensive,
and robust endotoxin bioassays of high impact in the biotechnology industry.
Terms: <Affinity><Algorithm Design><Algorithmic Design><Algorithmic Engineering><Assay><Binding><Bioassay><Biochemical><Biological><Biological Agent><Biological Assay><Biological Markers><Biological Products><Biosensor><Biotech><Biotechnology><Blood><Blood Reticuloendothelial System><COVID-19><CV-19><Cancers><Cardiac Diseases><Cardiac Disorders><Cell membrane><Cessation of life><Color><Complex Mixtures><Coronavirus Infectious Disease 2019><Cytoplasmic Membrane><Death><Detection><Development><Devices><Directed Molecular Evolution><Disease Marker><Dryness><Ecological impact><Endotoxins><Environmental Impact><Enzyme Gene><Enzymes><Esteroproteases><Evolution><Fever><Formulation><Future><Harvest><Health><Heart Diseases><High Throughput Assay><Horseshoe Crabs><Human><In Vitro><Industry><Injectable><International><Laboratories><Limulus><Lipopolysaccharides><Malignant Neoplasms><Malignant Tumor><Medical Device><Medicine><Methods><Modern Man><Molecular Interaction><Nature><Organ failure><Organism><Outcome><Peptidases><Peptide Hydrolases><Phagocytes><Phagocytic Cell><Pharmaceutical Agent><Pharmaceuticals><Pharmacologic Substance><Pharmacological Substance><Phase><Plasma Membrane><Predisposition><Property><Protease Gene><Proteases><Protein Engineering><Proteinases><Proteins><Proteolytic Enzymes><Publishing><Pyrexia><Reading><Reagent><Recombinant Proteins><Research Resources><Resources><Sampling><Septic Shock><Susceptibility><Sustainable Development><Techniques><Technology><Temperature><Testing><Therapeutic><Toxin><Variant><Variation><Xiphosura><algorithm engineering><algorithmic composition><amebocyte><bio-markers><biologic><biologic marker><biological sensor><biologics><biomarker><biopharmaceutical><biotherapeutic agent><coronavirus disease 2019><coronavirus disease-19><coronavirus infectious disease-19><cost><cost effective><design><designing><detection assay><detection limit><developmental><directed evolution><febrile><febris><genetic protein engineering><heart disorder><high throughput screening><improved><living system><luminescence><malignancy><manufacture><medical implant><neoplasm/cancer><novel><pharmaceutical><plasmalemma><protein design><response><safety testing><sensor><thermolability><thermostability><timeline>