A NEW METHOD FOR QUANTITATIVE DETERMINATION AND CHARACTERIZATION OF MICROVESICULES IN HUMAN ORGANISM LIQUIDS

摘要

1. A method for the quantitative and qualitative determination of exosomes in samples obtained from human cells or in body fluids, the method comprising the steps of:! a) capturing exosomes from a sample obtained from human cells or from body fluid with a primary anti-Rab 5b antibody; ! b) detecting exosome coupled antibodies for detection, said antibody for detection being an anti-CD63 antibody or an anti-caveolin-1 antibody; ! c) allowing the enzyme-associated secondary antibody to react with the antibody for detection; ! d) adding a substrate; and! e) reaction detection. ! 2. The method according to claim 1, where the method includes the step of purifying the exosome preparation from a sample obtained from human cells or body fluids, and stage a) is carried out on purified exosomes. ! 3. The method according to claim 1, where the body fluid is a sample of plasma, ascitic fluid, fluids of the brain, bone marrow, urine, feces or broncho-alveolar washout of a person. ! 4. The method according to claim 3, where the volume of body fluid used is less than 2 ml. ! 5. The method according to claim 1, where the body fluid or cell sample is obtained from a healthy individual and from a patient presumably having a tumor, and the detected number of exosomes in a patient’s sample is compared with the number of exosomes in a healthy individual’s sample, and an increase in the number is used as a sign of a tumor . ! 6. A non-invasive method for monitoring tumor growth, the method comprising the steps of:! a) periodic analysis of a sample of the patient’s body fluid; ! b) capture of exosomes from a sample obtained from human cells or from liquid