Monoclonal antibody for detecting mouse interleukin 6 as well as preparation method and application thereof

A monoclonal antibody and interleukin technology, applied in the field of immunoassay, can solve problems such as narrow linear range, low detection sensitivity, and radioactive contamination, and achieve the effects of wide linear range, good reaction specificity, and improved sensitivity and precision

Pending Publication Date: 2022-01-11
普健生物(武汉)科技有限公司
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  • Claims
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AI Technical Summary

Problems solved by technology

Radioimmunoassay has the disadvantages of short validity period and radioactive contamination
The ELISA method is usually only suitable for manual operation, which is not conducive to the detection of high-throughput automatic detection instruments, and increases the error caused by human factors in

Method used

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  • Monoclonal antibody for detecting mouse interleukin 6 as well as preparation method and application thereof
  • Monoclonal antibody for detecting mouse interleukin 6 as well as preparation method and application thereof
  • Monoclonal antibody for detecting mouse interleukin 6 as well as preparation method and application thereof

Examples

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Example Embodiment

[0059] Example 1

[0060] In this example, the monoclonal antibody of mouse interleukin 6 contains a first antibody and a second antibody, the first antibody secretes by hybridoma cell line 20-A4-D5, the second antibody from hybridoma cells The strain 10-B8-C9 secretes, the hybridoma cell lines 20-A4-D5 and the hybridoma cell lines 10-B8-C9 were prepared as follows:

[0061] S1, artificially synthetic mice interleight dielectric interleukin 6 gene, recombinantly group the mouse interleukin 6 gene into expression vector plasmid PATX1 to obtain an IL6-PATX1 expression vector, electrophoresis verification of the carrier, figure 1 Shown in figure 1 It can be seen that the cloned site is EcoRI / XHOI, the sequence of mouse interleukin 6 gene is shown in SEQ ID NO.13; the IL6-PATX1 expression vector is transfected into a 293F cell line for 6 days, collecting the supernatant The liquid was purified by nickel column to give mice interleukin 6 (IL6); purified IL6 for SDS-PAGE electrophores...

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Abstract

The invention provides a monoclonal antibody for detecting mouse interleukin 6 as well as a preparation method and application thereof, and belongs to the technical field of immunoassay. According to the invention, an artificially synthesized mouse interleukin 6 gene is subjected to plasmid recombination, protein expression, animal immunization and screening, and finally, a hybridoma cell strain 20-A4-D5 and a hybridoma cell strain 10-B8-C9 are obtained through subcloning; and an antibody secreted by the hybridoma cell strain 20-A4-D5 is labeled with acridinium ester to obtain a detection antibody, an antibody secreted by the hybridoma cell strain 10-B8-C9 is labeled with biotin to obtain a capture antibody, and the detection antibody and the capture antibody are applied to a chemiluminescence kit for detecting mouse interleukin 6. The kit provided by the invention is high in detection sensitivity, good in precision, wide in linear range and good in antigen reaction specificity, and does not generate cross reaction.

Description

technical field [0001] The invention relates to the technical field of immune analysis, in particular to chemiluminescence immunoassay technology, and in particular to a monoclonal antibody for detecting mouse interleukin-6 and a preparation method and application of the monoclonal antibody. Background technique [0002] Interleukin, also known as interleukin, mouse interleukin 6 (IL-6 or IL6) is a pleiotropic, α-helical, 22-28 kDa phosphorylated and variable glycosylated cytokine that plays a role in acute phase reactions, inflammation , hematopoiesis, bone metabolism and cancer progression. Together with tumor necrosis factor alpha (TNF-α) and interleukin 1 (IL-1), IL-6 drives the acute inflammatory response. IL-6 is almost entirely responsible for the febrile and acute phase responses of the liver and plays a very important role in the transition from acute inflammation to acquired immunity or chronic inflammatory disease. In addition to its functions in inflammation an...

Claims

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Application Information

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IPC IPC(8): C07K16/24G01N33/577G01N33/68G01N33/532
CPCC07K16/248G01N33/577G01N33/6869G01N33/532C07K2317/56C07K2317/51C07K2317/515G01N2333/5412Y02A50/30
Inventor 杨查理张永霞万定一鲁亮秦伏波代腾飞
Owner 普健生物(武汉)科技有限公司
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