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Human IL-15 subtype protein and preparation method and application thereof

A protein and sequence technology, applied in the field of human IL-15 subtype protein preparation, can solve problems such as limiting the application of IL-15 subtype protein and prone to allergic reactions

Active Publication Date: 2013-09-11
KUSN BEIRUIKANG BIOLOGICAL TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Because the mouse IL-15 subtype protein is immunogenic, it is easy to produce allergic reactions caused by anti-antibodies, which limits the application of IL-15 subtype protein, and human IL-15 subtype can overcome this defect

Method used

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  • Human IL-15 subtype protein and preparation method and application thereof
  • Human IL-15 subtype protein and preparation method and application thereof
  • Human IL-15 subtype protein and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0061] Example 1, preparation of purified functional untagged human IL-15 subtype protein

[0062] 1. Extraction of total cellular RNA

[0063] Take human peripheral blood cells, prepare peripheral blood mononuclear cell suspension, use 0.1 μg / ml LPS to act for 72 hours to stimulate peripheral blood mononuclear cell PBMC, obtain stimulated peripheral blood mononuclear cell suspension PBMC, add 1ml of lysing solution Trizol, repeat Mix by pipetting, dissolve the cells in Trizol, incubate the sample at room temperature for 5min to lyse the cells; add 0.2ml chloroform, mix vigorously for 15s, incubate the sample at room temperature for 3-5min, centrifuge at 12000g for 15min at 4°C; transfer the upper colorless aqueous phase after centrifugation To the EP tube, add 0.5ml of isopropanol, mix well, ice bath for 10min; centrifuge at 12000g for 10min at 4°C, carefully remove the supernatant; add 1ml of 75% ethanol, mix by inverting, centrifuge at 7500g for 5min at 4°C, wash twice Dis...

Embodiment 2

[0144] The concentration of LPS used in step 1 in this embodiment is 10 μg / ml, and the action time is 24 hours. The rest of the reagents and operating steps are exactly the same.

Embodiment 3

[0146] The concentration of LPS used in step 1 of this example is 20 μg / ml, and the action time is 24 hours; the rest of the reagents and operation steps are exactly the same.

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Abstract

The invention relates to preparation of human monoclonal antibody protein, in particular to preparation and an application of the human IL-15 subtype protein. In the invention, the human IL-15 subtype protein is prepared when an immune system is in an excited state; the antagonism of the IL-15 protein of a normal person is detected by an in-vitro test so as to provide the application of the subtype protein in clinical treatment on an autoimmune disease and the like. Studies prove that the subtype protein is not found in peripheral blood cells which are not stimulated by LPS, and the subtype protein is found in peripheral blood of the normal person stimulated by LPS, which show that the subtype protein possibly plays an important negative feedback regulation and control function under the condition of immune system activation. The subtype protein is found under the condition of immune system activation for the first time, and relevant research on the function is made, thus providing more direct evidence for developing the function in organic immune regulation and control.

Description

technical field [0001] The invention relates to the preparation of a human monoclonal antibody protein, in particular to a preparation method and application of a human IL-15 subtype protein. Background technique [0002] IL-15mRNA (interleukin 15, IL-15) is widely expressed in various tissues, including placenta, skeletal muscle, kidney, heart, lung, brain, intestinal epithelium, and also expressed in myeloid hematopoietic stem cells such as macrophages ,Dendritic Cells. IL-15 is structurally homologous to interleukin 2 (IL-2), and the IL-15 receptor consists of a high-affinity IL-15 receptor α chain, IL-2 / 15 receptor β chain and public chain γ chain, so IL-15 has some functions similar to IL-2, such as stimulating T cell activation and proliferation, enhancing NK cell killing activity and promoting B cells to produce immunoglobulin. Recent studies have found that IL-15 plays an important role in the differentiation, proliferation, and activation of NK cells, NKT cells, a...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/54C12N15/24C12N15/70A61K38/20A61P37/02A61P1/00A61P1/16A61P5/44A61P11/00A61P11/06A61P17/06A61P19/02A61P19/04A61P19/08A61P25/00A61P29/00
Inventor 刘海燕胡博
Owner KUSN BEIRUIKANG BIOLOGICAL TECH
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