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Anti-3 type mycoplasma urealytium MB protein antibody and immunochromatography kit applying antibody

Ureaplasma urealyticum and immunochromatography technology, applied in the field of biomedicine, can solve the problems of high production cost, low polyclonal antibody specificity, low titer and the like

Active Publication Date: 2016-09-28
HUBEI UNIV OF TECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The biggest advantage of monoclonal antibodies is high specificity, but the preparation method is cumbersome and the production cost is high, which limits its application
Polyclonal antibodies have defects such as low specificity and low titer

Method used

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  • Anti-3 type mycoplasma urealytium MB protein antibody and immunochromatography kit applying antibody
  • Anti-3 type mycoplasma urealytium MB protein antibody and immunochromatography kit applying antibody
  • Anti-3 type mycoplasma urealytium MB protein antibody and immunochromatography kit applying antibody

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0102] Example 1 Preparation of Rabbit Anti-Type 3 Ureaplasma Urealyticum MB Protein Monoepitope Antibody

[0103] The preparation method of rabbit anti-type 3 Ureaplasma urealyticum MB protein single epitope antibody is as follows:

[0104] 1) After structural biology analysis and related experimental research, 14 amino acids at positions 105-118 of the Ureaplasma urealyticum type 3 MB protein (GenBank sequence number AAC41437.1) were selected as the preparation of rabbit anti-Ureaplasma urealyticum type 3 MB protein antibody The linear antigenic epitope of this segment is KLPREPKPNEQLTI, and this sequence is named MB3Linear;

[0105] 2) After adding a cysteine ​​to the C-terminus of the amino acid sequence MB3Linear described in step 1), the polypeptide is synthesized and purified with an automatic polypeptide synthesizer, and the purified polypeptide is coupled with the carrier protein KLH to form an MB3Linear-KLH composite protein ;

[0106] 3) emulsifying the composite ...

Embodiment 2

[0112] Preparation of embodiment 2 anti-MB protein polyclonal antibody:

[0113] 1) Cloning of related genes

[0114] Bioinformatics analysis was performed on the MB protein of Ureaplasma urealyticum type 3 (the accession number in the NCBI protein database is AAC41437.1), and the peptide with the most abundant antigenic epitope in the N-terminal extracellular conserved domain was obtained, and the peptide was found At the same time, the 5' end of the DNA coding sequence was introduced with a restriction site NdeI, and the 3' end was introduced with a termination signal TAA and a restriction site XhoI. After that, the entire gene sequence was chemically synthesized respectively (full sequence synthesis was handed over to Completed by GenScript Biotechnology Co., Ltd., the artificially synthesized gene fragments were all connected to the vector pUC57 at the time of delivery), denoted as MB3, and the full sequence of the gene is shown in the sequence table. The protein sequence...

Embodiment 3

[0126] Example 3 Preparation and Application of Immunochromatography Kit Based on Quantum Dot Labeling Technology

[0127] 1. Quantum dot-labeled antibody AbMB3Linear

[0128] Add 0.4nmol carboxyl water-soluble quantum dots and 800nmol carbodiimide (EDC) to the microcentrifuge tube successively, make the volume to 1ml with MES buffer (10.66g / L MES, 0.74g / L EDTA pH 7.4), keep After 5 minutes of reacting at 37°C, 0.34 mg of the antibody AbMB3Linear prepared in Example 1 was added, and reacted in the dark for 2 hours. Adding single-terminal amino polyethylene glycol (PEG2000-NH2) to a final concentration of 1% (m / v), block the unreacted activated carboxyl site, and continue to react in the dark for 1h. The reacted sample was centrifuged with an ultrafiltration tube (molecular weight cut-off 100k), centrifuged at 6500g for 5min to a volume of 200ul, transferred the ultrafiltered sample to an ordinary EP tube, and centrifuged to remove aggregates (10000g, 3min). Add the supernat...

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Abstract

The invention relates to an anti-3 type mycoplasma urealytium MB protein antibody and an immunochromatography kit applying the antibody to detect the mycoplasma urealytium. The anti-3 type mycoplasma urealytium MB protein antibody is an antibody for identifying linear epitopes consisting of 105 to 118 amino acids of a 3 type mycoplasma urealytium MB protein; a serial number of the 3 type mycoplasma urealytium MB protein in GenBank is AAC41437.1; the sequence of 105 to 118 amino acids of the 3 type mycoplasma urealytium MB protein is KLPREPKPNEQLTI. The rabbit anti-3 type mycoplasma urealytium MB protein antibody provided by the invention has the characteristics of good specificity, high purity, high titer and low preparation cost.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and relates to an anti-type 3 Ureaplasma urealyticum MB protein antibody and an immunochromatographic kit using the antibody to detect Ureaplasma urealyticum. Background technique [0002] Ureaplasma urealyticum (M. urealyticum) is the only species in the genus Ureaplasma, named for its need for urea for growth. Ureaplasma urealyticum has two biological groups and 14 serotypes, which often exist in healthy asymptomatic people, and it is speculated that only certain subgroups or subgroups are pathogenic. Through the grouping research on the urogenital tract of different populations, it is found that most of the genitourinary tracts that inhabit and cause infection in humans are the first group, and type 3 is the most common. After infection with Ureaplasma urealyticum, most patients have no obvious symptoms. Therefore, it is difficult for patients to detect it, and it is easy for doctors to m...

Claims

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Application Information

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IPC IPC(8): C07K16/12G01N33/569G01N33/558
CPCC07K16/1253G01N33/558G01N33/56933G01N2333/30
Inventor 胡征董俊杨波
Owner HUBEI UNIV OF TECH
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