Method and a Kit To Detect Malignant Tumors and Provide a Prognosis
AssigneeHansabiomed OU
InventorsFrancesco Lozupone, Stefano Fais, Mariantonia Logozzi, Antonio Chiesi, Natasa Zarovni
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Abstract
A method and kit is provided to quantifying and qualifying exosomes in human cell derived samples or in body fluid based on expression of TM9-superfamily proteins on the exosomes. Furthermore, a method and a kit to diagnose malignant tumors is provided. The disclosure also provides a method to monitor tumor growth.
Claims17
A method to quantify and qualify tumor-related exosomes in human cell derived samples or in body fluid, said method having the steps comprising:
- a) optionally purifying an exosome preparation from the human cell derived sample or body fluid;
- b) capturing exosomes of the purified exosome preparation or the human cell derived sample or body fluid with a primary capturing antibody against a housekeeping protein present on exosomes, said primary capturing antibody being selected from the group consisting of:
- anti-tetraspanins, anti-annexins and anti-Rab-proteins;
- c) detecting tumor-related exosomes from the captured total exosomes with a detection antibody, said detection antibody being selected from the group consisting of antibodies against proteins belonging to Transmembrane-9 superfamily;
- d) allowing an enzyme linked secondary antibody react with the detection antibody;
- e) adding substrate;
- and
- f) detecting the reaction.
A method to quantify and qualify tumor-related exosomes in human cell derived samples or in body fluid, said method having the steps comprising:
- a) optionally purifying an exosome preparation from the human cell derived sample or body fluid;
- b) capturing exosomes of the purified exosome preparation or the human cell derived sample or body fluid with a primary capturing antibody, said primary capturing antibody being selected from the group consisting of:
- antiiTM9SF1-, antiTM9SF2-, antiTM9SF3- and antiTM9SF4-antibodies;
- c) detecting tumor-related exosomes from the captured exosomes with a detection antibody, said detection antibody being selected from the group consisting of anti-Rab5-, anti-CD63 and anti-Cav-1-antibodies;
- d) allowing an enzyme linked secondary antibody react with the detection antibody;
- e) adding substrate;
- and
- f) detecting the reaction.
A non-invasive method to monitor tumor growth, said method comprising the steps of:
- a) periodically taking a body fluid sample of a patient;
- b) optionally purifying an exosome preparation from the body fluid sample;
- c) capturing exosomes of the purified exosome preparation or the body fluid sample with a primary capturing antibody, said primary capturing antibody being selected from the group consisting of:
- antiiTM9SF1-, antiTM9SF2-, antiTM9SF3- and antiTM9SF4-antibodies;
- d) detecting tumor-related exosomes from the captured exosomes with a detection antibody, said detection antibody being selected from the group consisting of anti-Rab5-, anti-CD63 and anti-Cav-1-antibodies;
- e) allowing an enzyme linked secondary antibody react with the detection antibody;
- f) adding substrate;
- g) detecting the reaction, and
- h) drawing a correlation between quantity of detected exosomes and size and/or invasiveness of the tumor.
A test kit for quantifying and qualifying exosomes in human cell derived samples or in body fluid, said kit comprising:
- a) instructions to optionally purify an exosome preparation from the human cell derived sample or from body fluid;
- b) a primary antibody preparation for capturing exosomes of a purified exosome preparation or a human body fluid sample;
- c) a detection antibody preparation for detecting bound exosomes;
- d) an enzyme linked secondary antibody preparation for reaction with the detection antibody;
- e) a substrate for the enzyme;
- f) a positive control consisting of a standard exosome preparation from a human cancer cell line expressing TM9SF protein(s) of interest;
- and
- g) instructions to compare the reaction of the sample with the reaction of the positive control.
Dependencies are inferred from the claim text.