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
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[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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