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Preparation of multi-epitope thymidine kinase 1 (TK1) antibody and use of multi-epitope TK1 antibody for early tumor detection and risk early warning in mass physical examination screening

A multi-epitope, antibody technology, applied in the application of high specificity, early tumor detection and risk warning, the preparation of high-sensitivity multi-epitope TK1 antibody, to achieve the effect of stable activity, high sensitivity and high specificity

Active Publication Date: 2012-06-20
SHENZHEN HUARUI TONGKANG BIOTECHNOLOGICAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although this serum kit-enhanced luminescence immunoblot detection system is used in physical examination screening, the detection range is between 0.3-2pM, but at 0.1-1pM, the error is sometimes greater than 30%, and the routine physical examination serum TK1 of the population is used. The ROC value analysis was carried out with the serum TK1 of cancer patients. The area under the ROC value curve was 0.96, the specificity was 0.991, and the maximum likelihood method ratio (+) was 56.04, which proved that the specificity of the antibody was very good and could be used for population screening. However, the sensitivity is 0.51%, and it is necessary to further prepare antibodies with high detection sensitivity, specificity and stability to improve the detection system, so as to better improve the detection accuracy in the range of 0.1-1pM

Method used

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  • Preparation of multi-epitope thymidine kinase 1 (TK1) antibody and use of multi-epitope TK1 antibody for early tumor detection and risk early warning in mass physical examination screening
  • Preparation of multi-epitope thymidine kinase 1 (TK1) antibody and use of multi-epitope TK1 antibody for early tumor detection and risk early warning in mass physical examination screening
  • Preparation of multi-epitope thymidine kinase 1 (TK1) antibody and use of multi-epitope TK1 antibody for early tumor detection and risk early warning in mass physical examination screening

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Example 1: Selection of specific polyepitope antigens

[0047] Such as figure 1 As shown, the software predicts that TK1 has no signal peptide and no transmembrane structure. The diagram shows the distribution of 234 amino acids in the single-chain TK1 protein, indicating that the distribution of amino acids is relatively uniform. The map provides options: 1) From the perspective of antigenicity, it is better to select the peptide at the place where the blue curve peak is high; 2) According to the distribution of hydrophilic amino acids, select the hydrophilic region; 3) Select the negative value Aromatic amino acids with strong hydrophobicity.

example 2

[0048] Example 2: Select the antigen of high sensitivity, high specificity multi-site combined anti-human TK1-IgY antibody

[0049] Selection of antigens for the preparation of highly sensitive and highly specific multi-site combined anti-human TK1-IgY antibodies: Since the partial sequence of human cervical cancer TK1 has the same epitope as other proteins, the obtained antibodies may be different from other non-specific proteins There is cross-reaction and purification is difficult. According to the published and known TK1 sequence (protein library), select the special peptide exposed on the surface of the protein. According to the specificity of the polypeptide predicted by the software, the polypeptide fragments are listed in the following table. Antigen fragments with strong TK1 specificity were screened, and TK1 N-terminal 23 peptide, C-terminal 20 peptide and C-terminal 28 peptide were designed as haptens.

[0050]

Embodiment 3

[0051] Example 3: Screening for specific polyepitope antigens

[0052] Selection of specific multi-epitope antigen: according to the well-known spatial structure of TK1, select the antigenic determinant exposed on the surface of TK1 protein, screen the antigenic fragment with strong immunity of TK1 according to Example 2, screen the N-terminal 23 peptide of TK1, and the C-terminal The 20 peptide and the C-terminal 28 peptide are antigen fragments with strong immunity. The amino acid sequences of the three segments are as follows (see sequence listing): 1) N-terminal 23 peptide (3-25): CINLPTVLPGSPSKTRGQIQVIL, 2) C-terminal 20 peptide (206 -225): CPVPGKPGEAVAARKLFAPQ, 3) C-terminal 28 peptide (198-225): AGPDNKENCPVPGKPGEAVAARKLFAPQ.

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Abstract

The invention provides a high-specificity and high-sensitivity coordinated antibody against human thymidine kinase 1 (TK1) prepared from an antigenic determinant consisting of 23 peptides at an N terminal, 20 peptides at a C terminal and 28 peptides at the C terminal of the TK1 monomer from a human hela cell and the use of the detection and diagnosis system of the antibody in tumor diagnosis. The antigenic determinant comprises the following amino acid sequences: the sequence of the 23 peptides (3-25) at the N terminal:CINLPTVLPGSPSKTRGQIQVIL; the sequence of the 20 peptides(206-225) at the C terminal: CPVPGKPGEAVAARKLFAPQ; and the sequence of the 28 peptides (198-225) at the C terminal: AGPDNKENCPVPGKPGEAVAARKLFAPQ. The invention also provides a method for preparing the antibody by using the antigen. An antibody kit provided by the invention has the characteristics of high sensitivity, high specificity, low cost and the like; and the early tumor can be detected and pre-warned in mass physical examination screening by enhanced chemiluminescence dot blot assay, immuno-histochemistry and the detection kit.

Description

Technical field: [0001] The present invention relates to an antibody and its application, in particular to the preparation of a highly specific and highly sensitive multi-epitope TK1 antibody and its application in early tumor detection and risk warning in population screening. Background technique: [0002] Cancer is the killer of human beings. Although there are many imaging detection methods and treatment methods, such as: chemotherapy, endocrine therapy, radiotherapy and surgery, although great progress has been made, it is far from being perfect. With the possibility of recurrence and metastasis. Therefore, early detection, especially early detection and early treatment of precancerous lesions is the best gospel for the recovery of patients. [0003] Cancer is a chronic disease in which cells proliferate abnormally. The abnormal and uncontrolled proliferation of cancer cells is caused by one or more mutations in genes related to proliferation in normal cells. Although...

Claims

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Application Information

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IPC IPC(8): C07K16/40C07K16/02C12N9/12G01N33/577G01N33/574G01N33/573
Inventor 周际李劲斯文・斯库格艾伦・何
Owner SHENZHEN HUARUI TONGKANG BIOTECHNOLOGICAL
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