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ELISA detection kit for epidermal growth factor receptor III type mutant

A technology for detection kits and variable regions, applied in measuring devices, instruments, scientific instruments, etc., can solve the problems of cumbersome operation, difficult operation, high cost, etc., and achieve the effect of simple operation

Active Publication Date: 2017-11-21
GUIZHOU MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] At present, methods such as real-time fluorescent quantitative PCR, direct sequencing, dHPLC and PCR-RFLP are mainly used to detect the target in clinical practice. These detection methods are cumbersome, costly and difficult to operate. A simple and convenient method for detecting EGFRvⅢ is imminent

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  • ELISA detection kit for epidermal growth factor receptor III type mutant
  • ELISA detection kit for epidermal growth factor receptor III type mutant
  • ELISA detection kit for epidermal growth factor receptor III type mutant

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Embodiment Construction

[0028] The present invention will be further described below in conjunction with the accompanying drawings and embodiments, but not as any limitation to the present invention.

[0029] The present invention is implemented like this:

[0030] The anti-EGFRvIII monoclonal antibody 3D9 of the present invention, including the variable region amino acid sequences of its heavy chain and light chain, is shown in the sequence list SEQ ID NO: 7 and SEQ ID NO: 8; the heavy chain and light chain can be The base sequence of the variable region is shown in the sequence list SEQ ID NO: 9 and SEQ ID NO: 10.

[0031] The preparation process of monoclonal antibody 3D9 of the present invention is as follows:

[0032] 1 immunized animal

[0033] Fifteen healthy BALB / c female mice were divided into experimental group (1:1 mixture of antigen and adjuvant), adjuvant group (adjuvant only) and blank control group (normal saline). The time and site of immunization are shown in Table 1, 50 μl was in...

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Abstract

The invention discloses an ELISA detection kit for an epidermal growth factor receptor III type mutant. The key point is that a genetic engineering technique is adopted for acquiring an anti-EGFRvIII single-chain antibody and the anti-EGFRvIII single-chain antibody, biotin antigen, avidin-HRP and a developing fluid are formed into the main compositions of the kit. A completion method is used for establishing a detection method for EGFRvIII in cells, serum or tissue culture. The ELISA detection kit has the advantages of high specificity, high sensitivity and high accuracy. The kit also can be used for common scientific research or clinic tumor screening.

Description

technical field [0001] The invention relates to a method for rapidly detecting EGFRvIII, which is suitable for detecting EGFRvIII in serum, cell culture or tissue culture. Background technique [0002] EGFR is a member of the epidermal growth factor receptor family and a transmembrane glycoprotein with a molecular weight of 170kD. It is widely distributed on the surface of mammalian epithelial cells, fibroblasts, glial cells, keratinocytes and other cells, and is overexpressed in most tumors, and the degree of expression is closely related to the degree of differentiation and malignancy of the tumor. Studies have found that there are many mutants of EGFR in malignant tumors, among which the epidermal growth factor receptor type Ⅲ mutant (EGFRvⅢ) is the most common deletion mutant. EGFRvⅢ is caused by the in-frame deletion mutation of the extracellular region of EGFR. Compared with the complete molecular structure of EGFR, exons 2-7 of the extracellular region of EGFRvⅢ are ...

Claims

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Application Information

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IPC IPC(8): G01N33/558G01N33/577G01N33/535
CPCG01N33/535G01N33/558G01N33/577
Inventor 刘亭杨友辉刘香香吴琼薛维娜李勇军何彬
Owner GUIZHOU MEDICAL UNIV