Anti-human IL-1alpha monoclonal antibody and application thereof

A monoclonal antibody, L-1F12 technology, applied in the field of microbial engineering, can solve the problem of unclear expression mechanism of malignant tumor cells, achieve the effect of good passaging ability, high affinity, and cost reduction

Inactive Publication Date: 2013-04-03
SUZHOU UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Moreover, the expression mechanism of IL-1α in malignant tumor cells is still unclear.

Method used

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  • Anti-human IL-1alpha monoclonal antibody and application thereof
  • Anti-human IL-1alpha monoclonal antibody and application thereof
  • Anti-human IL-1alpha monoclonal antibody and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1: Preparation of mouse anti-human IL-1α protein monoclonal antibody

[0045] The prokaryotic expression product of IL-1α gene was used as the immunogen to immunize BALB / c mice to obtain B cells sensitized by the immunogen. The specific operation method:

[0046] 1. Preparation of monoclonal antibody immunogen

[0047] 1.1 Amplification of the complete open reading frame fragment of IL-1α gene

[0048] It is very difficult to obtain and purify IL-1α protein directly from tissues or cells to prepare antibodies. Therefore, it is an effective method to prepare antibodies through genetic engineering expression products. It is a prerequisite to obtain genetic engineering expression products to call the target gene.

[0049] Peripheral mononuclear lymphocytes (PBMC) were isolated from fresh human blood, and then subjected to total RNA extraction, RT-PCR amplification and construction of the recombinant transfer vector pGEM-T-IL-1α. The recombinant plasmid was sequenced to obtai...

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Abstract

The invention which belongs to the field of microbial engineering concretely relates to an anti-human IL-1alpha monoclonal antibody and an application thereof. The monoclonal antibody is generated by a hybridoma cell strain L, and preservation information of the hybridoma cell strain L comprises that: the preservation unit is China center for type culture collection (CCTCC); the address is Wuhan University in China; the preservation date is 07 / 01 / 2011; the preservation number is CCTCC NO: C201102; and the systematized name is hybridoma cell strain L-1F12. The high specificity and high titer mouse anti-human IL-1alpha monoclonal antibody allows the rapid flux detection on the IL-1alpha to be realized by adopting various means of indirect enzyme-linked immunosorbent assay (ELISA), immunocytoflurescence, immunofluorescence flow cytometer analysis and the like.

Description

Technical field [0001] The invention belongs to the field of microbial engineering, and specifically relates to an anti-human IL-α monoclonal antibody and a preparation method and application thereof. Background technique [0002] IL-1 (IL-1α and IL-1β) is a multifunctional cytokine. In 1972, Gery et al. discovered that the culture supernatant of human leukocytes contained a soluble substance, which was originally named lymphocyte activating factor or endogenous pyrogen, osteoclast activating factor, melanoma cell growth inhibitory factor, etc. In 1979, it was named IL-1 internationally. IL-1 can be synthesized and secreted by a variety of cells, such as monocytes, macrophages, and dendritic cells. [0003] The IL-1 gene of mouse and human is located on chromosome 2 and contains 7 exons. IL-1 has high homology among different species. At the amino acid level, the homology of IL-1α and IL-1β in different species is 60%-70% and 75%-78% respectively; but in the same species, the h...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/20C07K16/24G01N33/577C12R1/91
Inventor 刘海燕林丹丹刘春亮
Owner SUZHOU UNIV
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