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Monoclone antibody against HnRNP A2/B1 antigen, its preparation and use

A monoclonal antibody, antibody technology, applied in the biological field, can solve problems such as restricted expression

Inactive Publication Date: 2006-09-06
WOWU BIOTECH ZHEJIANG PROV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

HnRNP A2 / B1 is highly expressed in embryonic lung, but its expression in mature tissues is restricted, and its expression pattern in carcinoma is similar to that seen in embryonic lung growth, and this expression pattern is expressed at protein level and Signaling molecule levels are consistent, showing that HnRNP A2 / B1 may be a cancer growth protein (Montuenga LM et al. Am J Respir Cell Mol Biol, 1998; 19:554)

Method used

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  • Monoclone antibody against HnRNP A2/B1 antigen, its preparation and use
  • Monoclone antibody against HnRNP A2/B1 antigen, its preparation and use
  • Monoclone antibody against HnRNP A2/B1 antigen, its preparation and use

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Embodiment 1 obtains monoclonal antibody

[0038] 1) Immunize mice and obtain spleen cell stock solution

[0039] The human lung cancer cell line GLC82 was intraperitoneally injected with 20 million cells into Balb / C mice, and then injected with the same number of cells for booster immunization 4 times. 24 hours after the last injection, separate the spleen of the mouse and prepare the spleen cell stock solution: cut the abdominal cavity of the mouse in an ultra-clean bench, take out the spleen, and place it in a sieve in a plate containing 20ml of serum-free RPMI-1640 culture solution . Use two elbow droppers, one to fix the spleen, the other to scrape the spleen, squeeze out the splenocytes to make a splenocyte suspension, transfer it into a centrifuge tube (generally counting more than 200 million is better), and centrifuge at 1200rpm for 10 minutes. Aspirate the supernatant, add 10 ml of serum-free RPMI-1640 medium (Hyclone) to suspend the cells, and centrifuge at...

Embodiment 2

[0049] The identification of embodiment 2 monoclonal antibody type

[0050] The type and subtype of the monoclonal antibody obtained in Example 1 were identified.

[0051] The ImmunoPure® Monoclonal antibody isotyping kit from Pierce was used to detect the supernatant of the CGMCC0854 hybridoma cell culture, and it was found that the monoclonal antibody belonged to the IgG1 / κ chain type.

Embodiment 3

[0052] Example 3 Immunohistochemical Identification Test

[0053]Paraffin sections were dewaxed in xylene and dehydrated in alcohol. This was followed by incubation with 3% hydrogen peroxide-methanol for 20 minutes. Rinse with PBS buffer, then microwave repair in pH 6.0 citrate buffer, and rinse with PBS. The ascites to be tested was diluted with three dilution ratios of 1:100; 1:1000; 1:10000, and the diluted ascites was dropped on the slices. Incubate at 37°C for 1 hour. Anti-IgG polyperoxidase was added dropwise and incubated at 37°C for 20 minutes, then rinsed with PBS. Afterwards, DAB was used for color development, counterstained with hematoxylin, transparent, and neutral gum to seal the slides. Observe under a microscope and make a judgment on the result. Take photos of positive results. The results are shown in Figure 1. The results showed that "HnRNP A2 / B1" monoclonal antibody had obvious intracytoplasmic antigen distribution in cancer tissues, but had no speci...

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Abstract

A monoclonal antibody able to specifically bind with the determinant of antigen HnRNP A2 / B1 in human cancer cell and tissue kytoplasm, the nucleic acid molecule for coding the antibody, its preparing process and its application in diagnosing and / or treating human tumor are disclosed.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a monoclonal antibody, which has specific reactive ability to HnRNP A2 / B1 antigen in human cancer cells and tissue cytoplasm. Background technique [0002] With the development of modern industry, environmental pollution is increasing day by day, and the incidence of lung cancer is also increasing year by year. In developed countries such as Europe and the United States, the mortality rate of lung cancer has taken the first place in malignant tumors, and the incidence rate in my country has also increased significantly in the past 20 years. Statistical data shows: male lung cancer death rate increases by 120.93%, and female increases by 90.41%, and the increase in rural areas is more obvious than that in cities (Zhou Huaqing, editor-in-chief of lung cancer basic research and clinical treatment progress, 1st edition, Beijing: Science Press, 1999, 1-15 ). [0003] At present, there i...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K16/18C12N15/12C12N15/63C12N15/06A61K39/395A61P35/00G01N33/577
Inventor 胡赓熙
Owner WOWU BIOTECH ZHEJIANG PROV