Anti-prothymosin α polyclonal antibody and its preparation method and application

A technology of polyclonal antibody and thymosin, which is applied in peptide preparation methods, botany equipment and methods, biochemical equipment and methods, etc., can solve the problems of less than 50% positive rate, many sample collection times, and great influence , to achieve the effect of high specificity and accuracy, good sensitivity and low cost

Inactive Publication Date: 2011-12-28
XIAMEN UNIV
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Problems solved by technology

However, urine cytology examination is complicated to operate, and there are disadvantages such as many sample collection times, large volume, and long time-consuming.
The test results are greatly affected by subjective factors, and the positive rate is low. For patients with stage I and II, the positive rate is less than 50%.
[0010] At present, there is no report about the preparation of ProTα polyclonal antibody by this method.

Method used

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  • Anti-prothymosin α polyclonal antibody and its preparation method and application
  • Anti-prothymosin α polyclonal antibody and its preparation method and application
  • Anti-prothymosin α polyclonal antibody and its preparation method and application

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Embodiment Construction

[0054] The following embodiments will further illustrate the present invention in conjunction with the accompanying drawings.

[0055] The preparation method of the anti-thymosin pro-alpha polyclonal antibody comprises the following steps:

[0056] 1) Preparation of GST-ProTα fusion protein

[0057] (1) According to the gene sequence of ProTα, design the upstream primer P1 and the downstream primer P2, the upstream primer P1 and the downstream primer P2 are respectively:

[0058] Upstream primer P1: 5'-GCGGATCCATGTCAGACGCAGCCGTA-3',

[0059] Downstream primer P2: 5'-GCGAATCCCTAGTCATCCTCGTCGGTC-3',

[0060] The upstream primer P1 was introduced into the BamHI restriction site, and the downstream primer P2 was introduced into the EcoRI restriction site.

[0061] (2) Obtain the full-length gene cDNA of prothymosin α by RT-PCR method, purify the PCR product and connect it with pMD18-T Vector, name the recombinant vector TP, transform DH5α competent cells, and sequence the ident...

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Abstract

The anti-thymosin pro-alpha polyclonal antibody and its preparation method and application relate to an anti-thymosin pro-alpha polyclonal antibody. The anti-prothymosin α polyclonal antibody is a polyclonal anti-GST-ProTα antiserum obtained after immunizing New Zealand white rabbits with a fusion protein prepared by using GST-ProTα fusion expression. First prepare the fusion protein GST-ProTα, and then prepare the antibody. It can be used for immunoblotting and ELISA immunoassay of various samples and cells containing ProTα components, and tissue endogenous ProTα. Using the antibody to detect the ProTα level in urine can be used to diagnose urinary system tumors such as bladder cancer, kidney cancer and prostate cancer. The antibody has high titer and strong specificity. Various samples containing ProTα can be detected. Using prothymosin α in urine as a tumor marker for the diagnosis of urinary system cancer. The total process does not exceed 4 hours, and the required amount of samples is small.

Description

technical field [0001] The invention relates to an anti-thymosin pro-alpha polyclonal antibody, in particular to an anti-thymosin pro-alpha polyclonal antibody and its preparation method and application. Background technique [0002] Worldwide, the incidence of bladder cancer ranks ninth among malignant tumors, and it is the most common tumor in the urinary system. In the United States, the incidence of bladder cancer ranks fourth among male malignancies, after prostate cancer, lung cancer, and colon cancer, and ranks ninth among female malignancies. In my country, it ranks 7th in male malignant tumors and 10th in females, with an incidence rate of about 7.4 / 100,000. The degree of malignancy of bladder cancer is not very high, and the 5-year survival rate of early diagnosis and treatment is 93%, but if the diagnosis and treatment cannot be detected in time, the 5-year survival rate of advanced patients is only 6%, and the mortality rate is much higher than that of bladder c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/70C07K19/00G01N33/574C12N15/16C07K1/22C07K16/06C07K16/26
Inventor 周克夫杨彩霞韩伟吴汉洲高吉青王世媛张亭刘升发邢金春李伟张长弓
Owner XIAMEN UNIV
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