Preparation and application of a kind of human icf45 monoclonal antibody

A monoclonal antibody, F5H1D1H2 technology, applied in the preparation and application of human ICF45 monoclonal antibody, can solve the problems of cell proliferation and weakened adhesion ability

Active Publication Date: 2015-10-28
BEIJING NORMAL UNIVERSITY
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  • Description
  • Claims
  • Application Information

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Problems solved by technology

In the tumor cell model Hela, cell proliferation and attachment ability were found to be significantly weakened

Method used

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  • Preparation and application of a kind of human icf45 monoclonal antibody
  • Preparation and application of a kind of human icf45 monoclonal antibody
  • Preparation and application of a kind of human icf45 monoclonal antibody

Examples

Experimental program
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Effect test

Embodiment 1

[0029] Example 1. Preparation of monoclonal antibodies

[0030] First, the acquisition of hybridoma cell lines

[0031] 1. Preparation of ICF45 protein

[0032] The amino acid sequence of ICF45 protein is sequence 1 in the sequence listing, and the nucleotide sequence is sequence 2 in the sequence listing; the accession number of its full-length cDNA sequence on NCBI Genebank is AY463216.

[0033] Design specific primers with BamH I restriction sites at both ends:

[0034] Upstream primer 5'-CGC GGATCC TGGGGCGCCTGTAAA-3';

[0035] Downstream primer 5'-CGCGGATCCCGCAAAAGGGTCAG-5';

[0036] The nucleotide sequence of the artificially synthesized ICF45 protein (ie, sequence 1) was used as a template, and the above-mentioned upstream primer and downstream primer were used for PCR amplification, and the PCR product was obtained.

[0037] The PCR product was digested with BamHI, and the recovered digested product was the same as pGEX-PUC-3T (this plasmid was constructed from pG...

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Abstract

The invention discloses preparation and method of a human ICF (Intracellular Fluid) 45 monoclonal antibody, provides mus musculus hybridoma (mus musculus hybridoma) F5H1D1H2 with the storage number of CGMCC (China General Microbiological Culture Collection Center) No.8659, and also provides mus musculus hybridoma (mus musculus hybridoma) F5H1D1H2 with the storage number of CGMCC No.8659. The experience result shows that a mus musculus hybridoma (mus musculus hybridoma) F5H1D1H2 can be obtained by screening; and the mus musculus hybridoma (mus musculus hybridoma) F5H1D1H2 is subjected to biological characteristics identification in order to further research the influence of ICF45 at protein level on normal physiological process of the cell, and provide basis and build foundation for further study on the corresponding functional relation and function in diseases, tumor and allergic reaction, as well as prepare potential drug protein capable of inhibiting corresponding diseases.

Description

technical field [0001] The invention relates to the preparation and application of a monoclonal antibody of Interphase Cytoplasmic Foci Protein-45 (ICF45), in particular to the preparation and application of a human ICF45 monoclonal antibody. Background technique [0002] ICF45 protein is a new cytoplasmic protein discovered, its expression is highly cell cycle phase specific and plays an important role in cell cycle regulation. Previous studies have shown that ICF45 localizes to the cytoplasm during interphase, forms 1-2 specific punctate foci near the nuclear membrane, colocalizes specifically with the centrosome in the cell cycle phase, and colocalizes with the centrosome during mitosis. disappear. [0003] Numerous studies suggest that tumorigenesis is related to centrosome expansion. Abnormal centrosome duplication, such as increased number of centrosomes, can cause multi-level spindles and abnormal chromosome segregation during mitosis. Stable silencing of ICF45 res...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/20C07K16/00G01N33/68G01N33/577C12R1/91
Inventor 何大澄商瑜伊力亚尔王珊珊肖雪媛
Owner BEIJING NORMAL UNIVERSITY
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