Preparation method of human TDP-43 monoclonal antibody coating ELISA plate and ELISA detection kit

Abstract

The invention relates to a preparation method of a new human TDP-43 monoclonal antibody coating ELISA plate and ELISA detection kit. The key technology mainly lies in that: genetic engineering method is utilized to prepare a specific mouse anti-human monoclonal antibody, and the ELISA plate by which the antibody is purified and then coated is taken as important component of the kit, and then the ELISA plate, TDP-43 polyclonal antibody and horseradish peroxidase labelled second antibody, TDP-43 standard substance, TDP-43 positive control, sample diluent, cleaning solution, development solution and stop solution are assembled into a humanized TDP-43 ELISA detection kit; a sensitive and rapid method for detecting concentration of TDP-43 in ELISA antibody capturing human serum or cell and tissue culture is established, and the method has high specificity and high stability. The kit is mainly used in general libratory or neuropathology clinical research.

Description

technical field [0001] The present invention relates to a new biological agent—a monoclonal antibody against human TDP-43, and an ELISA detection kit for human TDP-43, suitable for TDP-43 in human tissue, cell culture or serum quantitative detection. technical background [0002] TDP-43 (TAR DNA-binding protein) is the TAR DNA-binding protein, which is widely expressed in the nucleus of human cells, binds DNA and RNA, regulates the transcription and splicing of intracellular nucleic acids, and is also involved in the synthesis of small RNAs in the nucleus, apoptosis and cellular Split. After being discovered for the first time in 1995, the abnormal expression of TDP-43 has been detected in large numbers in the serum and autopsy of dementia patients. Studies have shown that TDP-43's intracellular expression, localization, protein length, mutation or hyperphosphorylation, and ubiquitination modification can all cause the loss or disorder of some brain tissue functions (Neuma...

AI Technical Summary

Problems solved by technology

[0010] At present, several major ELISA kit manufacturers in the world only produce TDP-43 protein or antibody alone, and there is no report on the production of a complete set of TDP-43 ELISA kits. The research on TDP-43 usually uses TDP-43 overexpression or wrong expression. Models of sense mutations, such as nerve cells, chicken embryos or mice, but lack of quantitative research on TDP-43 in human blood, tissues or cells

At the same time, the few products launched by domestic manufacturers are in the initial stage, and even a positive control and internal reference cannot be provided, so the measured results are naturally suspicious

On the one hand, the reason for the problem may be that the manufacturer’s quality awareness is indifferent; on the other hand, the purity and activity of the capture antibody and detection antibody may also be technically difficult to meet the corresponding requirements of the kit