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Cancer kit containing monoclonal antibody of specific binding ROS1

A monoclonal antibody, species-specific technology, applied in the direction of antibodies, specific peptides, anti-tumor drugs, etc., can solve the problem of no relevant reports on monoclonal antibodies, achieve inhibition of tumor cell growth, good binding specificity, and good application prospects Effect

Active Publication Date: 2020-07-24
GUANGDONG CELL BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] At present, the detection or treatment drugs developed for ROS1 are already on the market, but there is no relevant report on the development of monoclonal antibodies against the specific domain of ROS1

Method used

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  • Cancer kit containing monoclonal antibody of specific binding ROS1
  • Cancer kit containing monoclonal antibody of specific binding ROS1
  • Cancer kit containing monoclonal antibody of specific binding ROS1

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] The preparation of embodiment 1 immunogen

[0028] According to the structural properties of ROS1 protein, through analysis, the optimized immune polypeptide sequence ggdlltylrkarmatfygplltlvdlvdlcvdiskgcvylermhfihrdlaarnclvsvkdytsprivkigdfglardiykndyyrkrgegllpvrwmapeslmdgifttqsdvwsfgil was selected, and the corresponding amino acid sequence was obtained by peptide synthesis. The obtained protein was subjected to SDS-PAGE electrophoresis, and the results were as follows: figure 1 shown. The target protein is produced at the position of lane 1, and lane 2 is the blank control.

Embodiment 2

[0029] Example 2 Preparation of Hybridoma Cells for Monoclonal Antibody Production

[0030] 1. Animal immunity

[0031] After mixing the purified polypeptide of Example 1 and the immune enhancer PEI (1:3), it was dissolved in serum-free DMEM solution without double antibody, so that the final injection concentration was 1 mg / ml. 100 μl / mouse intramuscularly injected into 35-day-old mice, and injected once every 10 days. After the third immunization, local blood was collected from the mice, and the mouse serum was separated by centrifugation at 40C 10,000r / min for 20 minutes. After the serum was diluted 500-16,000 times, the serum antibody level was detected by indirect ELISA method. As a control, a group of blank experiments was designed at the same time, and the test results of the mouse with the best effect are shown in Table 1.

[0032] Table 1 Serum test results

[0033] Dilution factor OD value 500 1.936 1000 1.705 2000 1.556 4000 1.407 ...

Embodiment 3

[0043] Example 3 Preparation of Monoclonal Antibody Ascites

[0044] Prepare 50-56-day-old mice and inject 850 μl of paraffin oil sterilized by autoclaving (peritoneal injection).

[0045] After 6-9 days, hybridoma cells ROS1-1 were continued to be injected into the mice that had been injected with paraffin oil. After the recovered hybridoma cells were blown up and diluted with 1ml RPMI 1640 cell culture medium, the mice were injected intraperitoneally, and the physical state of the mice should be observed and recorded every day. When the abdomen swells to a certain extent, the mouse ascites should be extracted in time with a 5ml sterile syringe.

[0046] In the procedure for extracting ascites, the mouse's body was fixed first, and then the abdomen was disinfected with alcohol. Ascites was extracted by injecting a syringe into the mouse abdomen. After the ascites was collected, the second mouth continued to observe the situation. If the mouse's abdomen swells again, ascit...

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Abstract

The present invention relates to a cancer kit containing a monoclonal antibody of specific binding ROS1. The monoclonal antibody is obtained by taking an ROS1 kinase domain obtained by screening as anantigen and immunizing a mouse. The antibody has the relatively good binding specificity, and it is verified that the antibody has the function of inhibiting the growth of tumor cells, so that the kit has the relatively good application prospect.

Description

technical field [0001] The present invention relates to the field of biology, and more specifically relates to a cancer kit comprising a monoclonal antibody specifically binding to ROS1. Background technique [0002] ROS1 is an oncogene sequence that can be fused with the gag gene, and has certain tumorigenicity after fusion. The wild-type ROS1 gene is located in the q12 region of chromosome 6. It encodes 2347 amino acids with a molecular weight of 259kDa. The encoded protein is a transmembrane tyrosine kinase receptor protein, which consists of three parts: tyrosine kinase region, transmembrane region and extracellular region containing glycosylation site. The N-terminal 1861 amino acids constitute the extracellular region, the transmembrane region consists of about 1882 amino acids, and the C-terminal 464 amino acids constitute the cytoplasmic region. The membrane region consists of three YWTD domains folded into three β-helical structures interspersed in nine type III ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/32C07K16/40C07K14/82C12N9/12C12N15/13A61K39/395A61P35/00G01N33/574G01N33/573G01N33/577
CPCA61K2039/505A61P35/00C07K14/82C07K16/32C07K16/40C07K2317/92C12N9/12C12Y207/10001G01N33/573G01N33/574G01N33/577G01N2333/82G01N2333/912
Inventor 彭菲顾超
Owner GUANGDONG CELL BIOTECHNOLOGY CO LTD
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