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Biomacromolecule capable of binding to β-amyloid protein and its encoding dna, β-amyloid protein detection kit and use thereof

A technology of amyloid protein and detection kit, applied in the field of molecular biology, can solve the problem that the exact mechanism of AD pathogenesis is not completely clear, and achieve the effect of easy grasp and popularization, high sensitivity and quick response

Active Publication Date: 2019-03-01
ZHEJIANG JUKANG BIOENG CO LTD
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  • Description
  • Claims
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Problems solved by technology

So far, the exact mechanism of AD pathogenesis is not fully understood

Method used

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  • Biomacromolecule capable of binding to β-amyloid protein and its encoding dna, β-amyloid protein detection kit and use thereof
  • Biomacromolecule capable of binding to β-amyloid protein and its encoding dna, β-amyloid protein detection kit and use thereof
  • Biomacromolecule capable of binding to β-amyloid protein and its encoding dna, β-amyloid protein detection kit and use thereof

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Experimental program
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Embodiment

[0026] Embodiment: The technical route of the present invention is, first, use the antigen 43 peptide β-amyloid protein to immunize mice, after confirming that the immunization is successful, isolate the spleen and lymph nodes of the mice, extract the total RNA, reverse transcribe it into cDNA, and pass PCR The light and heavy chain genes of the antibody are amplified. Then the scFv DNA was assembled and amplified to construct the pCANTAB5E-scFv recombinant plasmid. The plasmid was transformed again to obtain a single-chain antibody phage library, and the target antibody was obtained through screening. Finally, an adult β-amyloid detection kit was assembled.

[0027] Construction and screening of single-chain antibodies by phage display technology

[0028] Cells and cell lines use: E. coli TG1, used for the transformation of pCANTAB-5E; E. coli HB2151, for scFv expression.

[0029] Trizol total RNA extraction kit, M-MLV reverse transcriptase, and RNase inhibitors were...

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Abstract

The invention relates to a biomacromolecule capable of binding to beta amyloid protein, a DNA encoding the macromolecule, a beta amyloid protein detection kit and uses thereof. Firstly, the mice were immunized with the antigen 43 peptide amyloid beta protein. After confirming the success of the immunization, the spleen and lymph nodes of the mice were isolated, the total RNA was extracted, reverse-transcribed into cDNA, and the light and heavy chain genes of the antibody were amplified by PCR; then Assemble and amplify the scFv DNA to construct the pCANTAB5E‑scFv recombinant plasmid; then transform the plasmid to obtain a single-chain antibody phage library, and obtain the target antibody through screening; finally assemble the adult β-amyloid protein detection kit. The antibody screened by the invention has a wide detection range of 0-1000pg / ml, a high linear correlation coefficient, a detection sensitivity of up to 3.0pg / ml, and is very stable.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to a biomacromolecule capable of binding to beta-amyloid protein, DNA encoding the macromolecule, a beta-amyloid protein detection kit and uses thereof. Background technique [0002] Population aging is a major social problem facing all countries in the world today, and has attracted widespread attention from the international community. By the end of 2014, my country's elderly population over 60 years old had reached 212 million, accounting for 15.5% of the total population. It is expected to double to 400 million around 2033, and by around 2050, the elderly population will reach one-third of the country's population. The "silver boom" will have a profound impact on my country's economic, social, political, and cultural development. The mental and physical health problems of the elderly have increasingly become the focus of attention of the whole society. Alzheimer's di...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68G01N33/543C07K16/18C12N15/13
CPCG01N33/54306G01N33/68G01N33/6896C07K16/18G01N2333/4709
Inventor 陶新博张守涛郭亚楠
Owner ZHEJIANG JUKANG BIOENG CO LTD