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Monoclonal antibodies capable of specifically recognizing mouse protein Doppel

A monoclonal antibody, specific technology, applied in the field of biomedicine to achieve the effect of improving reactogenicity, speeding up the preparation process, and improving the immune effect

Inactive Publication Date: 2018-07-27
FOURTH MILITARY MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since there is currently no specific antibody against mouse Doppel protein, the preparation of related antibodies is of great significance for in-depth research on the function of mouse and human Doppel

Method used

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  • Monoclonal antibodies capable of specifically recognizing mouse protein Doppel
  • Monoclonal antibodies capable of specifically recognizing mouse protein Doppel
  • Monoclonal antibodies capable of specifically recognizing mouse protein Doppel

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Embodiment Construction

[0037] The invention discloses a mouse-derived monoclonal antibody method for mouse Doppel protein: the mouse Doppel protein expressed by prokaryotic cells is used to couple carrier protein to make an immunogen, and after immunization by dot membrane and subcutaneous embedding method, 2 antibodies are screened out. Strain anti-mouse Doppel protein monoclonal antibody hybridoma cell line. The hybridoma cell line has the ability to stably secrete antibodies. The antibody prepared and purified from ascites can specifically recognize the mouse Doppel protein, and finally obtained the gene sequence of its heavy chain variable region, confirming the uniqueness of its corresponding protein sequence. It provides a powerful research tool for in-depth study of the functions of mouse, human and rat Doppel proteins.

[0038] The present invention will be described in detail below in conjunction with the preparation method of specific monoclonal antibodies, antibody activity detection, cha...

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Abstract

The invention discloses monoclonal antibodies capable of specifically recognizing the mouse protein Doppel, belonging to the technical field of biomedicine. The monoclonal antibodies can bind to the protein Doppel expressed in natural mouse brains and prokaryotic cells and specifically recognize amino acid residues at position 82-90 of the protein Doppel, wherein the amino acid sequence of the amino acid residues at position 82-90 is as shown in SEQ ID No. 1. According to the invention, the heavy-chain variable region genes of two monoclonal antibodies, i.e., 1A9 and 1D6, are cloned, the genesequences of heavy-chain variable regions and amino acid sequences are obtained, and the uniqueness of the gene sequences and protein sequences are confirmed. Since the recognized epitope sequence NYWQFPDGI is completely identical in the protein Doppel in mice, rats and human beings, it is indicated that the two monoclonal antibodies have critical application value in research on the functions ofthe protein Doppel.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and relates to a heavy chain variable region of a monoclonal antibody specifically recognizing mouse Doppel protein and a gene thereof. Background technique [0002] Mouse Doppel protein was discovered in 1999, because its gene is located at the 16kb downstream of the mouse normal prion protein gene (Prnp), and its structure and function are similar to those of mice and human prion protein (Prion), so the original description of Prnp do wnstream p rion-like p Doppel is named after rotein (Doppel means "double, double" in German, abbreviated as Dpl, and translated into Chinese as "folded protein"). [0003] The sequence of the mouse Doppel protein gene (Prnd) is relatively conservative, and it has greater homology with the genes of fish, tetrapods, cattle, sheep and humans. Its function involves important physiological functions such as neuron development and reproduction, but specific, Exh...

Claims

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Application Information

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IPC IPC(8): C07K16/18C12N15/13
CPCC07K16/18C07K2317/565C07K2317/92
Inventor 丁天兵王媛高欢
Owner FOURTH MILITARY MEDICAL UNIVERSITY
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