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AssigneeCodiak BioSciences, Inc.
InventorsDamian J. HOUDE, Young Jun CHOI, John D. KULMAN, Douglas E. WILLIAMS
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Described herein are novel rapid and reliable methods of detection of extracellular vesicles and quantifying extracellular vesicle concentrations and absolute number from various sources, including raw cell harvest. The methods described herein comprise detection of intrinsic fluorescence of extracellular vesicles in biological samples. Extracellular vesicles analyzed by the methods of this application have a stereotypical elution profile distinct from known contaminants. The methods described herein are a significant improvement over the state of the art and fulfills an unmet need in the field of extracellular vesicle manufacturing and quality control.
A method of detecting extracellular vesicles, comprising:
The method of any one of the preceding claims wherein said intrinsic fluorescence emission signal is determined at an excitation wavelength of 550 nm and an emission of 570 nm.
The method any one of the preceding claims, wherein said sample is separated into fractions prior to determining said intrinsic fluorescence emission signal.
The method of any one of the preceding claims, wherein said sample is subjected to a filtration step prior to determining said intrinsic fluorescence emission signal.
The method of any one of the preceding claims, wherein said sample is subjected to a centrifugation separation step prior to determining said intrinsic fluorescence emission signal.
The method of any one of the preceding claims, wherein said sample is subjected to a sucrose density gradient step prior to determining said intrinsic fluorescence emission signal.
The method of any one of the preceding claims, wherein said sample is a subjected to a separation step comprising use of a density gradient prior to determining said intrinsic fluorescence emission signal.
The method of any one of the preceding claims, wherein said sample is derived from a cell culture.
The method of any one of the preceding claims, wherein said sample is derived from a body fluid of an animal.
The method of any one of the preceding claims, comprising determining an amount of said extracellular vesicles within said sample comprising comparing said intrinsic fluorescence emission signal to the intrinsic fluorescence emission signal of a standard.
The method of any one of the preceding claims, comprising quantifying the amount of said extracellular vesicles within said sample, based upon the area under of the curve of a chromatogram comprising said intrinsic fluorescence emission signal.
The method of any one of the preceding claims, comprising determining an amount of said extracellular vesicles within said sample comprising comparing said intrinsic fluorescence emission signal to the luminescence signal of a standard.
The method of any one of the preceding claims, wherein said extracellular vesicle is an exosome.
The method of any one of the preceding claims, wherein said extracellular vesicle comprises a therapeutic payload.
A method of detecting extracellular vesicles, comprising:
Dependencies are inferred from the claim text.