US-12625148-B2 - Method for collecting extracellular vesicles derived from nervous system cells

Abstract

The present invention addresses a problem of providing a method for collecting extracellular vesicles derived from nervous system cells at an improved efficiency. This problem is solved by a method for collecting extracellular vesicles derived from nervous system cells, said method comprising a step for mixing an anti-APLP1 antibody with a sample containing extracellular vesicles to form anti-APLP1 antibody-extracellular vesicle complexes and a step for collecting the anti-APLP1 antibody-extracellular vesicle complexes.

Inventors

• Takashi Kudo
• Kanta YANAGIDA
• Shoshin AKAMINE

Assignees

• OSAKA UNIVERSITY
• TOSOH CORPORATION

Dates

Publication Date

20260512

Application Date

20201223

Priority Date

20191227

Claims (4)

1. 1 . A method for collecting extracellular vesicles derived from nervous system cells comprising: mixing an anti-amyloid beta precursor like protein 1 (anti-APLP1) antibody and a sample containing extracellular vesicles derived from nervous system cells to form a complex of the anti-APLP1 antibody and the extracellular vesicle, and collecting the complex of the anti-APLP1 antibody and the extracellular vesicle, thereby collecting the extracellular vesicles derived from the nervous system cells.
2. 2 . The method for collecting the extracellular vesicles according to claim 1 , wherein the sample is obtained by crudely purifying the extracellular vesicles from a specimen containing the extracellular vesicles derived from nervous system cells before mixing the anti-APLP1 antibody and the sample.
3. 3 . The method for collecting the extracellular vesicles according to claim 2 , wherein the extracellular vesicles are crudely purified from the specimen by a size exclusion chromatography, an ultracentrifugation, an affinity purification, a polymer precipitation, or a combination thereof.
4. 4 . The method according to claim 1 , wherein the sample is obtained from a specimen selected from the group consisting of whole blood, serum, plasma, lymph, urine, ascites, pleural effusion, cerebrospinal fluid, intercellular fluid, tears, nasal discharge, and saliva, and the specimen contains the extracellular vesicles derived from the nervous system cells.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS This application is a U.S. National Stage application of PCT/JP2020/048209 filed 23 Dec. 2020, which claims priority to Japanese Application No. 2019-239839 filed 27 Dec. 2019, the entire disclosures of which are hereby incorporated by reference in their entireties. TECHNICAL FIELD The present description discloses a method for collecting extracellular vesicles derived from nervous system cells, a method for detecting a neuropsychiatric disorder, a method for collecting components derived from nervous system cells, a reagent for collecting extracellular vesicles, and a kit for detecting extracellular vesicles containing the reagent. BACKGROUND ART Patent Literature 1 discloses biomarkers and diagnostic and prognostic methods for Alzheimer's disease and other neurodegenerative disorders. Patent Literature 2 discloses that biomarkers in vesicles (for example, exosomes) isolated from a biological sample are detected to be used for diagnosis and prognostication of Alzheimer's disease and other neurodegenerative disorders. Patent Literature 3 discloses methods for quantifying subpopulations of exosomes and diagnostic and prognostic methods for neurodegenerative disorders (for example, Alzheimer's disease). Patent Literature 4 discloses how to use exosomes and exosomal biomarkers in diagnostic and prognostic methods for neurological disorders, immunological disorders, placental diseases, cancer, hematological disorders, kidney disease, gastrointestinal diseases, liver diseases, and musculoskeletal diseases. CITATION LIST Patent Literature Patent Literature 1: WO2015/061634Patent Literature 2: WO2017/193115Patent Literature 3: WO2018/094120Patent Literature 4: WO2019/144056 SUMMARY OF INVENTION Technical Problem Patent Literatures 2 to 4 disclose use of exosomes for diagnosis and prognostication of neurodegenerative disorders. However, for example, NCAM or CD171 used for collecting exosomes described in Patent Literature 2 is not a protein specific to nervous system cells. In order to efficiently collect extracellular vesicles derived from nervous system cells, it is necessary to collect extracellular vesicles derived from nervous system cells using a protein highly specific to other nerve cells. The present invention addresses a problem of providing a method for collecting extracellular vesicles derived from nervous system cells at an improved efficiency. Solution to Problem As a result of diligent research, the present inventors have found that extracellular vesicles derived from nervous system cells can be efficiently collected by performing immunoprecipitation targeting APLP1 present in the extracellular vesicles. The present invention includes, for example, the following aspects as an embodiment. Item 1. A method for collecting extracellular vesicles derived from nervous system cells comprising a step of mixing an anti-APLP1 antibody and a sample containing extracellular vesicles to form a complex of the anti-APLP1 antibody and the extracellular vesicle, and a step of collecting the complex of the anti-APLP1 antibody and the extracellular vesicle. Item 2. The method for collecting the extracellular vesicles according to item 1, wherein the sample containing the extracellular vesicles contains the extracellular vesicles crudely purified from a specimen. Item 3. The method for collecting the extracellular vesicles according to item 1 or 2, wherein the crude purification is performed by a size exclusion chromatography, an ultracentrifugation, an affinity purification, a polymer precipitation, or a combination thereof. Item 4. A method for detecting the neuropsychiatric disorder comprising a step of obtaining measured values of polypeptides and/or polynucleotides that are a biomarker of the neuropsychiatric disorder from the extracellular vesicles collected by the method according to any one of claims 1 to 3, and a step of comparing the measured value with a corresponding reference value to determine whether the measured value is within or outside a reference range. Item 5. The method for detecting the neuropsychiatric disorder according to item 4, wherein the neuropsychiatric disorder is selected from neurodegenerative disorder, post-cerebral spinal trauma neurologic dysfunction, brain tumor, infection-related cerebrospinal disorder, multiple sclerosis, schizophrenia, and bipolar disorder. Item 6. The method for detecting the neuropsychiatric disorder according to item 5, wherein the neurodegenerative disorder is selected from dementia, Parkinson's disease, amyotrophic lateral sclerosis, progressive supranuclear palsy, multiple system atrophy, and triplet repeat disease. Item 7. A method for collecting components derived from the nervous system cells comprising a step of collecting at least one biomolecule selected from the group consisting of sugar, lipid, polypeptide, and polynucleotide from the extracellular vesicles collected by the method according to any one of items