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Ffa Sciences LLC

Alan Marc Kleinfeld

Kleinfeld, Alan M.

Andrew Henry Huber

James Patrick Kampf

Baolong Zhu

Thomas Kwan

Identification and use of proteins fluorescently labeled and that undergo a change in fluorescence index upon binding bilirubin are described. Probes are disclosed which are labeled at a cysteine or lysine residue and also probes labeled at both cysteine and lysine with two different fluorophores. These probes are useful for determination of unbound bilirubin levels in a fluid sample.

Development and use of fluorescent probes of unbound bilirubin

The invention relates to determination of the profile of unbound free fatty acids (FFAu) in biological samples, such as human and animal blood specimens and plant and animal oils, by measuring the fluorescence response of sets of different fluorescently labeled fatty acid binding proteins (probes) that undergo a change in fluorescence ratio at 2 wavelengths upon binding an FFAu. Use of these profiles in human and animal disease, in basic research, in drug development and in industrial uses is disclosed.

Determination of the unbound free fatty acid profiles in biological and industrial specimens

A method for high throughput discovery of proteins fluorescently labeled at a cysteine residue and that undergo a change in fluorescence ratio at 2 wavelengths upon binding an unbound analyte is described. Probes are disclosed which are labeled at a cysteine residue and also probes labeled at both cysteine and lysine with two different fluorophores. These probes are useful for characterization and measurement of hydrophobic species in a fluid sample, particularly characterization and measurement of levels of unbound free fatty acids. A profile of unbound free fatty acids can be determined for an individual which can be used to determine the individual's relative risk for disease.

Development and use of cysteine-labeled fluorescent probes of unbound analytes

Methods of determining levels of unbound metabolites are disclosed. Probes derived from fatty acid binding protein muteins are described that bind preferentially to a number of unbound metabolites including oleate, stearate, linoleate, palmitate, arachidonate and unconjugated bilirubin. A profile for a patient is determined using one or more of the described probes. The profile is useful in diagnosis of disease, particularly myocardial infarction, non-alcoholic fatty liver disease (NAFLD), diabetes, stroke, sepsis and neonatal jaundice. The responses of multiple probes to a test sample are used to classify the degree of acute coronary syndrome by comparison to multi-probe profiles generated from unstable angina, non ST elevation myocardial infarction, and ST elevation myocardial infarction.

Use of probes for unbound metabolites

A method for high throughput screening of probes is described. These probes are useful for characterization and measurement of unbound metabolites in a fluid sample, particularly characterization and measurement of levels of unbound free fatty acids. By practice of the disclosed invention, a profile of unbound metabolites can be determined for an individual which can be used to determine the individual's relative risk for disease such as stroke, cardiac disease and cancer.

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Encinitas, CA, United States of America

Andrew Henry Huber