Method for producing insoluble aggregate of neurodegenerative-disease-related protein

摘要

The purpose of the present invention is to develop a method for amplifying in vitro to a large amount of a homogenous insoluble aggregate that is equivalent to an insoluble aggregate formed in the brain of a patient. A method of producing an insoluble aggregate including TDP-43 protein and fragments thereof according to the present invention includes the steps of: (1) introducing an insoluble fraction originated from the brain of a neurodegenerative disease patient into a cell culture in which the intact TDP-43 protein can be expressed in a constitutive manner; (2) culturing the cultured cell into which the insoluble fraction has been introduced; and (3) separating an insoluble fraction from the cultured cell. Optionally, the method may additionally include a step of amplifying the insoluble aggregate of the neurodegenerative-disease-related protein in the cultured cell.

主权项

1. A method of amplifying in vitro a homogenous insoluble aggregate comprising TAR DNA-binding protein 43 kDa (TDP-43) protein and fragments thereof, comprising:
(1) a step of introducing an insoluble aggregate originated from a brain of a neurodegenerative disease patient or from a biological sample of said patient into a cultured cell expressing a full-length TDP-43 protein in a constitutive manner; (2) a step of incubating the cultured cell with the insoluble aggregate already introduced thereinto, to obtain an amplified insoluble aggregate; and (3) a step of separating an amplified insoluble aggregate from the incubated cultured cell.