Cancer diagnosing kit containing HLA-G monoclonal antibodies and use thereof

A monoclonal antibody, kit technology, applied in biological testing, material testing products, measuring devices, etc., to achieve the effect of high sensitivity

Active Publication Date: 2009-02-04
四川首创生物制药有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0015] For the existing HLA-G monoclonal antibody as a cancer detection marker and the defect of HLA-G enzyme-linked immunoassay, the purpose of the present invention is to provide a new monoclonal antibody, i.e. anti-HLA-G monoclonal antibody, named It is HGY-2, which is produced by the hybridoma cell line with the preservation number CCTCC NO.C200722 preserved by the China Type Culture Collection Center, and belongs to the IgG1 subtype

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  • Cancer diagnosing kit containing HLA-G monoclonal antibodies and use thereof
  • Cancer diagnosing kit containing HLA-G monoclonal antibodies and use thereof
  • Cancer diagnosing kit containing HLA-G monoclonal antibodies and use thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0040] Example 1 Preparation of HGY monoclonal antibody of the present invention

[0041] According to the method disclosed by Ye Shangmian patent: ZL 2004 10040919.X, the anti-HLA-G monoclonal antibody HGY was prepared using the hybridoma cell line with the preservation number CCTCC NO.C200416 preserved in the China Center for Type Culture Collection. The purification of the natural HLA-G protein, the preparation of the monoclonal antibody and the characteristics of the monoclonal antibody are all consistent with the content disclosed in the above patent.

[0042] Preparation of HGY-2 monoclonal antibody of the present invention

[0043] According to Ye Shangmian's patent: the method disclosed in ZL 2004 10040919.X, the basis of the present invention also adopts the preparation method of HGY monoclonal antibody, using the natural HLA-G protein purified from placental tissue aborted within three months of pregnancy, wherein Containing all HLA-G transcript isomers and glyc...

Embodiment 2

[0053] Example 2 Development of HLA-G enzyme-linked immunoassay kit of the present invention

[0054] 1. Monoclonal antibody:

[0055] The kit uses two anti-HLA-G protein molecule-specific monoclonal antibodies (HGY and HGY-2). These two monoclonal antibodies can bind to different antigenic epitopes of HLA-G protein molecules. Antibodies were stored at -80°C.

[0056] 2. Preparation of HLA-G protein standard:

[0057] The HLA-G protein standard used in the kit is obtained from the collected induced abortions by immunoaffinity chromatography using an anti-HLA-G monoclonal antibody (4H84 that has been commercialized or Ye Shangmian’s patent: HGY in ZL 2004 10040919). It is obtained by separating and purifying the extract of stage 1 placenta tissue, and its purity is greater than 95%. The method of gene recombination can also be used to clone and establish a cell line expressing HLA-G protein, and then use anti-HLA-G antibody immunoaffinity chromatography to separate and pu...

Embodiment 3

[0073] Example 3 Characteristics of the HLA-G enzyme-linked immunoassay kit of the present invention

[0074] The present invention first establishes a sandwich type enzyme-linked immunoassay kit for detecting plasma / serum soluble HLA-G, and verifies the specificity, sensitivity, accuracy and repeatability of the kit.

[0075] ● image 3 A standard curve of 10 HLA-G ELISA assays is shown. The results show that the Sandwich ELISA kit established by the present invention has a sensitivity of 10 ng / ml and high repeatability (the average intra-assay and inter-assay errors of each standard point are less than 10%; Table 2). In each batch of testing, three doses of positive control samples were set up for the HLA-GELISA. It can also be seen from Table 2 that the intra-assay and inter-assay errors of the positive control samples are both less than 10%; further confirming the reproducibility of the kit.

[0076] Table 2. Reproducibility of HLA-G enzyme-linked immunohistochemical ...

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Abstract

The present invention discloses a cancer diagnostic kit containing the anti-HLA-G monoclonal antibody, and an application thereof. The kit is an enzyme linked immunosorbent cancer diagnostic kit which is prepared with the novel anti-HLA-G monoclonal antibody and the anti-HLA-G monoclonal antibody HGY-2; wherein, the HGY-2 is generated with the hybridoma which is conserved in the Chinese Typical Culture Conservation Center and is numbered as CCTCC NO.C200722. The kit adopts the serum or other body fluid as the detected sample, the operation is convenient, the accuracy rate is high, the patient compliance is excellent, and the kit provides a novel method and means for cancer diagnosis.

Description

technical field [0001] The present invention relates to a monoclonal antibody, in particular to an anti-human leukocyte antigen G (HLA-G) monoclonal antibody and a hybridoma cell line secreting the antibody. [0002] The invention also relates to an enzyme-linked immunoassay kit for diagnosing malignant tumors prepared by using the antibody, and the clinical application of the kit in cancer diagnosis, prognosis evaluation, selection of treatment plan and treatment monitoring. Background technique [0003] Human leukocyte antigen G (HLA-G) is a nonclassical class I leukocyte antigen (see Geraghty et al. Proc Natl Acad Sci US A. 1987; 84:9145-9). The gene of the leukocyte antigen is located on the short arm of the sixth pair of chromosomes; the gene and the expressed product are 86% identical to the classical class I leukocyte antigen (HLA-A, B and C). Compared with the classic class I leukocyte antigens, HLA-G has two important features: First, under normal physiological con...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/52G01N33/577
Inventor 叶尚勉
Owner 四川首创生物制药有限公司
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