Primer, cDNA, vector, AQP4 monoclonal antibody, and preparation method of AQP4 monoclonal antibody

Abstract

The invention belongs to the technical field of DNA recombination, and discloses a primer, a coded cDNA, a vector, an AQP4 monoclonal antibody, and a preparation method of the AQP4 monoclonal antibody. A cho-cell clone expressing rat AQP4 m23 is established; the cDNA of the encoded rat AQP4 m23 is coned from total RNA extracted from a rat brain through using a primer reverse transcription polymerase chain reaction; and the cDNA is inserted into NheI and SacII sites of a perres 2-egfp vector after sequencing is carried out. The anti-mouse AQP4 and the anti-human AQP4 prepared in the invention have high affinity to AQP4 in the central nervous system in mice, and are proved to induce neuromyelitis optica lesions and symptoms in mice in vitro and in vivo. The studying and new treatment methodexploration of the neuromyelitis optica are benefited.

Description

technical field [0001] The invention belongs to the technical field of DNA recombination, and in particular relates to a primer, an encoded cDNA, a carrier, an AQP4 monoclonal antibody and a preparation method. Background technique [0002] At present, the existing technologies commonly used in the industry are as follows: Antigen mouse in vivo immunization method: use ready-made antigens or synthetic polypeptides as antigens, inject them into mice, and obtain antibodies by inducing immune responses in mice, and finally carry out antibody purification. Phage antibody technology: clone all variable region genes of a certain animal on the surface of phage for display and expression, and use different antigens to screen clones carrying specific antibody genes to obtain specific antibodies. [0003] To sum up, the problems existing in the prior art are: [0004] (1) According to the literature, the antigen immunization method cannot induce the production of effective antibod...

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Problems solved by technology

[0004] (1) According to the literature, the effective antibody against aquaporin 4 (anti-AQP4) cannot be induced in mice by antigen immunization, that is, the produced antibody has no pathogenic effect, so it cannot be applied to the establishment of a model of neuromyelitis optica

[0005] (2) Since the antigens required by phage antibody technology can only be obtained by peptide synthesis and do not have the spatial folding structure of proteins, the screened antibodies cannot bind well to receptors in the body, so the affinity and virulence significantly reduced disease

[0006] (3) Anti-aquaporin 4 antibodies (anti-AQP4 antibodies) currently on the market are all used to detect aquaporin 4 (AQP4), but they do not have the effect of causing disease in animals, so they cannot be used in disease models the establishment of

[0011] It is still very difficult to establish an animal model of neuromyelitis optica

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