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Antigenic polypeptide of tumor suppressor p16 and its application

A technology of tumor suppressors and antigenic peptides, applied in the field of biological immunology, can solve the problems of low sensitivity and specificity, high cost, etc., and achieve the effects of improving detection efficiency, reducing economic costs, and improving sensitivity

Active Publication Date: 2020-11-06
SHENZHEN HUAZHONG BIOLOGICAL PHARMA MACHINERY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The purpose of the present invention is to overcome the above-mentioned deficiencies of the prior art, to provide a tumor suppressor p16 antigenic polypeptide and a test kit containing the tumor suppressor p16 antigenic polypeptide for detecting p16 autoantibody levels in blood samples, to solve the current situation The sensitivity and specificity of the method of using autoantibodies as tumor diagnostic markers is not high, and the cost is high

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  • Antigenic polypeptide of tumor suppressor p16 and its application
  • Antigenic polypeptide of tumor suppressor p16 and its application
  • Antigenic polypeptide of tumor suppressor p16 and its application

Examples

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Embodiment 1

[0065] This embodiment 1 provides an antigenic polypeptide of tumor suppressor p16, said antigenic polypeptide of tumor suppressor p16 contains the amino acid sequence shown in SEQ ID NO:1. Of course, the amino acid sequence with the same function obtained by deleting, inserting or replacing the amino acid sequence shown in SEQ ID NO: 1 is also within the scope disclosed in this embodiment.

[0066] The synthesis method of the antigen polypeptide of the tumor suppressor p16 provided in Example 1 is as follows:

[0067] 1. Synthesize the gene sequence encoding polypeptide 1 through the amino acid sequence of polypeptide 1, add His-tag tags before and after the gene sequence of polypeptide 1, and insert a digestive enzyme cleavage site between the gene sequence encoding polypeptide 1 and the His-tag tag point.

[0068] 2. DH5α plasmid amplification vector transformation:

[0069] 1. Activate the bacteria inserted into the polypeptide 1 gene sequence overnight at 37°C;

[0070...

Embodiment 2

[0120] This embodiment provides a method for combining the antigenic polypeptide of the tumor suppressor p16 (abbreviated as P16 antigenic polypeptide) with serum and plasma IgG in Example 1, the method is as follows:

[0121] Coat the P16 polypeptide antigen with a 96-well microplate plate, set the concentration gradient at 1.0 μg to 5.0 μg / ml, set up double wells, bind the primary antibody with QC, and finally obtain the SBI specific binding curve by microplate reader detection.

[0122] Wherein, the SBI binding curve is as follows Figure 4 shown. Depend on Figure 4 It can be seen that when the P16 concentration is 1.0-4.0 μg / ml, the SBI value gradually decreases with the increase of the concentration; when the P16 antigen polypeptide concentration is 4.0-4.5 μg / ml, the SBI value gradually increases with the increase of the concentration. The SBI binding curve shows that when the P16 antigen polypeptide is at a lower concentration of 1.0 μg / ml, the bottom of the 96-well ...

Embodiment 3

[0124] Example 3 provides a kit for detecting p16 autoantibody levels in blood samples containing the antigenic polypeptide of the tumor suppressor p16 in Example 1.

[0125] 1. The kit for detecting the p16 autoantibody level in a blood sample provided in this example comprises the reagents described in Tables 1-8 below:

[0126] Table 1. Antigen Coating Buffers

[0127]

[0128] Table 2. Working antigens

[0129]

[0130]

[0131] Table 3. Reference antigens

[0132]

[0133] Table 4. 0.1M PBST wash buffer (1000ml volume)

[0134]

[0135] Table 5. Sample Dilution Assay Solution

[0136]

[0137] Table 6. Secondary Antibody Standard Solution

[0138]

[0139] Table 7. Stop Buffers

[0140]

[0141] Table 8. Substrate chromogenic solution

[0142]

[0143] 2. The operation method of the kit for detecting p16 autoantibody levels in blood samples in this embodiment is as follows:

[0144] (1) Coating: Dilute the working antigen and reference anti...

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Abstract

The invention discloses an antigen polypeptide of a tumor inhibition factor p16, and an application thereof. The polypeptide active fragment of the antigen polypeptide of the tumor inhibition factor p16 is at least one of an amino acid sequence (1) represented by SEQ ID NO: 1 and an amino acid sequence (2) obtained through deleting, inserting or replacing the amino acid sequence (1) represented by SEQ ID NO: 1 and having the same functions. The antigen polypeptide can be applied to the preparation of a kit for detecting the p16 autoantibody level in a blood sample. The antigen polypeptide has a specific antigenic determinant, so the specific binding rate of the antigen polypeptide and the autoantibody of the tumor inhibition factor p16 contained in a sample to be tested is improved, and the inspection sensitivity and the detection efficiency are increased, thereby the risk of early tumors can be accurately predicated, and reliable data is provided for the study of new tumor drugs.

Description

technical field [0001] The invention belongs to the technical field of biological immunology, and relates to an antigenic polypeptide of tumor suppressor p16 and its application and a kit for detecting p16 autoantibody level in blood samples containing the antigenic polypeptide of tumor suppressor p16. Background technique [0002] A large number of studies have shown that tumor-associated antigens in serum or plasma can induce the body to produce autoantibodies. In the first 3-5 years of malignant tumor transformation, high concentrations of tumor-associated antigen autoantibodies can appear in the blood of patients. This shows that both antibodies can be used to detect tumor antigens, and tumor antigens can also be used to detect autoantibodies, but the specificity and sensitivity of using tumor autoantibodies to detect tumors are much higher than using tumor antigens to detect tumors. The reason is that many tumor-associated antigens exist not only in tumor patients, but ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/47G01N33/68G01N33/574
CPCC07K14/4703G01N33/57488G01N33/68G01N2333/4704G01N2800/50G01N2800/52
Inventor 肖承荣易经达
Owner SHENZHEN HUAZHONG BIOLOGICAL PHARMA MACHINERY
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