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Preparation of multi-epitope TK1 antibody and application of multi-epitope TK1 antibody to evaluation on recurrence risk and prognosis of tumor patient at early stage

A multi-epitope, antibody technology, applied in the direction of anti-enzyme immunoglobulin, material test, immunoglobulin from egg, etc., to achieve high sensitivity, high yield and high purity.

Active Publication Date: 2012-05-02
SHENZHEN HUARUI TONGKANG BIOTECHNOLOGICAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the detection range of this serum kit-enhanced luminescence immunoblotting detection system is between 0.3-2pM in the trial and physical examination screening, the error is sometimes greater than 30% when the detection range is 0.1-1pM

Method used

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  • Preparation of multi-epitope TK1 antibody and application of multi-epitope TK1 antibody to evaluation on recurrence risk and prognosis of tumor patient at early stage
  • Preparation of multi-epitope TK1 antibody and application of multi-epitope TK1 antibody to evaluation on recurrence risk and prognosis of tumor patient at early stage
  • Preparation of multi-epitope TK1 antibody and application of multi-epitope TK1 antibody to evaluation on recurrence risk and prognosis of tumor patient at early stage

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Example 1: Selection of specific polyepitope antigens

[0046] 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

[0047] Example 2: Select the antigen of high sensitivity, high specificity multi-site combined anti-human TK1-IgY antibody

[0048] 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.

[0049]

[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 (325): 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 coordination combination anti-human TK1 antibody prepared from an antigenic determinant consisting of 23 peptide at an N end, 20 peptide at a C end and 28 peptide at the C end of human cervical cancer cells, and application thereof to tumor diagnosis. The antigenic determinant comprises the following amino acid sequences: 1) the 23 peptide (3-25) at the N end: CINLPTVLPGSPSKTRGQIQVIL: 2) the 20 peptide (206-225) at the C end: CPVPGKPGEAVAARKLFAPQ; and 3) the 28 peptide at the C end: AGPDNKENCPVPGKPGEAVAARKLFAPQ. The invention simultaneously provides a method applying the antigen for preparing the antibody. An antibody kit provided by the invention has the characteristics of high sensitivity, high specificity, low cost and the like, tumor rehabilitation populations are periodically monitored by an enhanced chemiluminescence dot blot detection method, immunohistochemistry detection and the detection kit, and the recurrence and transfer risk and the prognosis of the tumor rehabilitation people are evaluated.

Description

Technical field: [0001] The present invention relates to an antibody and its application, specifically the preparation of a highly specific and highly sensitive multi-epitope TK1 antibody and its application in the assessment of early recurrence risk and prognosis of tumor patients. 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. There is a possibility of recurrence and metastasis, among which the five-year recurrence or metastasis rate of tumor patients is as high as 40%, and the mortality rate of relapsed patients is as high as 80%. [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 ce...

Claims

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Application Information

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IPC IPC(8): C07K16/40C07K16/02C12N9/12G01N33/53G01N33/574
Inventor 周际李劲斯文・斯库格艾伦・何
Owner SHENZHEN HUARUI TONGKANG BIOTECHNOLOGICAL
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