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Preparation method of thymidine kinase 1 antibody and application of thymidine kinase 1 antibody in proliferation of ureaplasma urealyticum

A technology of Ureaplasma urealyticum and thymidine kinase, which is applied in anti-enzyme immunoglobulins, transferases, measuring devices, etc., can solve problems such as drug resistance of Ureaplasma urealyticum, unqualified reagents, double infection, etc., and achieve stable activity, Good stability and high purity

Active Publication Date: 2015-06-03
SHENZHEN HUARUI TONGKANG BIOTECHNOLOGICAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The reasons are very complex, such as resistance of Ureaplasma urealyticum, detection error or unqualified reagents, superinfection (people who have used antibiotics for a long time are prone to fungal and other insensitive bacterial infections)

Method used

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  • Preparation method of thymidine kinase 1 antibody and application of thymidine kinase 1 antibody in proliferation of ureaplasma urealyticum
  • Preparation method of thymidine kinase 1 antibody and application of thymidine kinase 1 antibody in proliferation of ureaplasma urealyticum
  • Preparation method of thymidine kinase 1 antibody and application of thymidine kinase 1 antibody in proliferation of ureaplasma urealyticum

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Experimental program
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Effect test

Embodiment 1

[0032] Ureaplasma thymidine kinase 1 (UuTK1 ), known as a tetramer, is structurally similar to hTK1 . The monomer is symmetrically formed by four 223 amino acids, and the C-terminal helix of UuTK1 interacts with one helix of the adjacent monomer. Due to genetic differences, only 30% of the sequence of the identified hTK1-related structure is very similar to Uu-TK, and the sequences of TK1 at the N and C terminals are very different. The three-dimensional crystal structure analysis of UuTK1 provides the basis for the preferred active peptide of the present invention as an immune antigen. The present invention relates to UuTK1 antibodies that recognize the entire UuTK1 protein. The entire UuTK1 protein specifically recognized by the antibody provided by the invention and any polypeptide with epitope activity include: UuTK1 protein, monomeric, 2- or 4-meric TK1 protein with enzymatic activity / inactivity, or TK1 protein and other molecules Formed protein complexes or inhibitors....

Embodiment 2

[0035] Embodiment 2 immune antigen preparation

[0036] The chemical synthesis obtained 10 amino acid residues as a hapten and 96 amino acid residues as UuTK1 as a standard quality control product. The purity was greater than 96% as identified by HPLC, and the quality was qualified by mass spectrometry. Then amidate the C-terminal of the hapten 10 peptide, and connect cysteine ​​at the N-terminal (entrusted to Shenzhen Hanyu Company, China)

[0037] CRHHHKVPNRPYLNSNSEEFIKFFKN-amidation

[0038] The above-mentioned optimized hapten was cross-linked with BSA to form 10-peptide-BSA immune antigen (chemical synthesis entrusted to Shenzhen Hanyu Company, China).

Embodiment 3

[0040] UuTK1 antibody was prepared according to the existing mono / polyclonal antibody technology with the immunization antigen of Embodiment 2. Implementation 3 is the step of preparing the anti-TK1 IgY crude product according to the hen immunization procedure and the IgY antibody screening technology, taking the IgY antibody screening technology as an example. For detailed steps, see Examples 5, 6 and 7 in Patent ZL02134727.1, "Preparation of Anticytoplasmic Thymidine Kinase-IgY and Composition for Tumor Diagnosis".

[0041] The hens were immunized with the above-mentioned 10-peptide-BSA immunization antigen and antibodies were prepared. The advantages of using hen immunization are:

[0042] (1) There are molecular genetic differences between chicken IgY and human IgG;

[0043] (2) There are racial differences between chicken TK1 and human TK1;

[0044] (3) Compared with the polyclonal antibody prepared by traditional rabbit immunization, the IgY extracted from the egg yol...

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Abstract

The invention provides design of an antigen for ureaplasma urealyticum thymidine kinase 1 (Ureaplasma urealyticum thymidine kinase1, UuTK1) and a preparation method of an antibody for the antigen. The antibody of UuTK1 and a composition kit of the antibody are capable of specifically identifying a UuTK1 antigen and used for detecting the proliferation degree of the ureaplasma urealyticum. The antibody kit has the characteristics of high sensitivity, high specificity, low cost and the like and is used for being applied to detecting the proliferation speed of the ureaplasma urealyticum in people with human urogenital tract and newborn respiratory tract infection diseases.

Description

【Technical field】 [0001] The present invention relates to a method for preparing an antibody and its application, in particular to a method for preparing an antibody against Ureaplasma urealyticum thymidine kinase 1 (UuTK1) and its use in detecting human urogenital and respiratory tract infections Proliferation of Ureaplasma urealyticum. 【Background technique】 [0002] Ureaplasma urealyticum (Uu for short) is a prokaryotic cell without a cell wall isolated from the urethral secretions of patients with nongonococcal urethritis (NGU). , which includes 2 biogroups, 14 serotypes. The biological group 1 is called Ureaplasma parvum (Up for short), containing 4 serotypes 1, 3, 6, and 14, while the biological group 2 continues to be called Ureaplasma urealyticum (Uu for short), containing serum Types 2, 4, 5, 7, and 7 to 13. Currently, reports on the association of serotypes 1 and 3 of biogroup 1 and serotypes 4 and 8 of biogroup 2 with infection and pathogenicity are controversia...

Claims

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Application Information

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IPC IPC(8): C07K16/40C12N9/12G01N33/569
Inventor 周际艾伦·何斯文·斯库格郭林李远
Owner SHENZHEN HUARUI TONGKANG BIOTECHNOLOGICAL
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