Fusion protein of IL2 and single chain antibody against GD2, its encoding genes and use thereof

A single-chain antibody, fusion protein technology, applied in applications, genetic engineering, plant genetic improvement, etc., can solve problems such as shortness of breath, thrombocytopenia, shock and coma, and achieve low side effects, good effects, and good biological activity. Effect

Inactive Publication Date: 2006-01-18
SHENYANG INST OF APPLIED ECOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Although IL-2 has an independent anti-tumor effect, repeated application of IL-2 to the human body can produce a variety of side effects, mil

Method used

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  • Fusion protein of IL2 and single chain antibody against GD2, its encoding genes and use thereof
  • Fusion protein of IL2 and single chain antibody against GD2, its encoding genes and use thereof
  • Fusion protein of IL2 and single chain antibody against GD2, its encoding genes and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] A kind of IL2 whose rare codon is changed by mutation has the base sequence in SEQ ID No: 1 in the sequence table (see Chinese patent application number: 03111487.3 for details).

[0023] GCTCCGACTTCTAGCTCTACCAAGAAAACCCAGCTGCAGCTGGAACA

[0024] CCTGCTGCTGGATTTACAGATGATTCTGAACGGTATCAACAACTACAA

[0025] GAACCCGAAACTGACCCGTATGCTGACCTTCAAGTTTTACATGCCGAA

[0026] GAAAGCTACCGAACTGAAACATTCTTCAGTGTCTAGAAAGAAGAACTCA

[0027] AACCTCTGGAGGAAGTTCTGAACCTGGCTCAGAGCAAAAACTTTCAC

[0028] CTGCGTCCGCGTGACTTAATCAGCAATATCAACGTAATAGTTCTGGAA

[0029] CTGAAAGGTTCTGAAACCACCTTCATGTGCGAATATGCTGATGAGACA

[0030] GCAACCATTGTAGAATTTCTGAACCGTTGGATTACCTTCTCTCAGTCTA

[0031] TCATTCCTACTCTGACT

[0032] (1) Information of SEQ ID No: 1 (see sequence listing)

[0033] (a) Sequential features

[0034] * Length: 399 bp

[0035] * Type: nucleic acid

[0036] * Chain type: double chain

[0037] * Topology: Linear

[0038] (b) Molecular type: cDNA

[0039] (c) Assumption: No

[0040] (d) Anti...

Embodiment 2

[0050] A mouse-derived anti-GD2 single-chain antibody gene M-ScFv is derived from hybridoma HB8759 and has the base sequence of SEQ ID No: 2 in the sequence table.

[0051] CAGGTGAAGCTGCAGGAGTCAGGAGGAGGCTTGGTGCAACCTGGAG

[0052] GATCCATGAAACTTCTCCTGTGTTGTCTCTGGATTCACTTTCAGTAATTA

[0053] CTGGATGAACTGGGTCCGCCAGTCTCCAGAGAAGGGGCTTGAGTGGA

[0054] TTGCAGAAATTAGATTGAAATCCAATAATTTTGCAAGATATTATGCGGA

[0055] GTCTGTGAAAGGGAGGTTCACCATCTCAAGAGATGATTCCAAAGGTA

[0056] GTGTCTACCTGCAAATGATCAACCTAAGAGCTGAAGATACTGGCATTT

[0057] ATTACTGTACCAGTTATGGTAACTACGTTGGGCACTATTTTGACCACTG

[0058] GGGCCAAGGGACCACGGTCACCGTCTCCTCAGGTGGAGGCGGTTCAG

[0059] GCGGAGGTGGCTCTGGCGGTGGCGGATCGGACATTGAGCTCACCCAG

[0060] TCTCCAAAATTCATGTCCACATCAGTAGGAGACAGGGTCACAGGTCACC

[0061] TGCAAGGCCAGTCAGAATGTGGATACTAATGTAGCCTGGTATCAACAA

[0062] AAACCAGGGCAATCTCCTGAACCACTGCTTTTCTCGGCATCCTACCGT

[0063] TACACTGGAGTCCCTGATCGCTTCACAGGCAGTGGATCTGGGACAGAT

[0064] TTCACTCTCACCATCAGCAATGTGCAGTCTGAAGACTTGGCAGAGTAT

[0065...

Embodiment 3

[0085] The IL2-M-ScFv fusion gene has the base sequence of SEQ ID No: 3 in the sequence table:

[0086] ATGGCTCCGACTTCTAGCTCTACCAAGAAAACCCAGCTGCAGCTGGA

[0087] ACACCTGCTGCTGGATTTACAGATGATTCTGAACGGTATCAACAACTA

[0088] CAAGAACCCGAAACTGACCCGTATGCTGACCTTCAAGTTTTACATGCC

[0089] GAAGAAAGCTACCGAACTGAAACATTCTTCAGTGTCTAGAAGAAGAAC

[0090] TCAAACCTCTGGAGGAAGTTCTGAACCTGGCTCAGAGCAAAAAACTTT

[0091] CACCTGCGTCCGCGTGACTTAATCAGCAATATCAACGTAATAGTTCTG

[0092] GAACTGAAAGGTTCTGAAACCACCTTCATGTGCGAATATGCTGATGAG

[0093] ACAGCAACCATTGTAGAATTTCTGAACCGTTGGATTACCTTCTCTCAG

[0094] TCTATCATCTCTACTCTGACTGGTGGAGGCGGTTCACAGGTGAAGCTG

[0095] CAGGAGTCAGGAGGGGGCTTGGTGCAACCTGsAGGATCCATGAAACT

[0096] CTCCTGTGTTGTCTCTGGATTCACTTTTCAGTAATTACTGGATGAACTGG

[0097] GTCCGCCAGTCTCCAGAGAAGGGGCTTGAGTGGATTGCAGAAATTAG

[0098] ATTGAAATCCAATAATTTTGCAAGATATTATGCGGAGTCTGTGAAAGG

[0099] GAGGTTCACCATCTCAAGAGATGATTCC AAAGTAGTGTCTACCTGCA

[0100] AATGATCAACCTAAGAGCTGAAGATACTGGCATTTATTACTGTAGCAG

[0101] TTATGG...

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Abstract

The present invention relates to biomedicine preparation, and is especially the expression in colibacillus and application of one new type of IL-2-antiGD2 single strand antibody fusion gene. IL-2 with rare codon altered in mutation is connected with single strand antibody of melanoma to form the fusion protein with antitumor effect. The fusion protein has the amino acid sequence as shown in SEQ ID No. 4 in the sequence list, and can activate intracorporeal immune system to kill specifically tumor cells of melanoma, fibroblast tumor, small cell lung carcinoma, etc. Compared with IL-2, the fusion protein has less side effects, even high bioactivity and even high tumor cell killing effect.

Description

technical field [0001] The invention relates to biopharmaceuticals, in particular to the expression and application of a novel IL2-anti-GD2 single-chain antibody fusion gene in Escherichia coli. Background technique [0002] Biological response modifiers (biological response modifiers, BRMs) mainly refer to some molecules and immune cells from the organism itself. Such factors are broad and include cytokines, interleukin 2 (Il-2), interferon (INF), tumor necrosis factor (TNF), mAb receptors, LAK cells, anti-idiotypic antibody vaccines and molecular vaccines Wait. BRM alone regulates the immune ability of the body and kills tumor cells, and has achieved good curative effect. However, since only part of it reaches the tumor target site after it is injected into the body, its killing effect cannot be fully exerted, and it has toxic and side effects. The BRM is coupled to the monoclonal antibody, and the monoclonal antibody carries it to the target site to play a role in kill...

Claims

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Application Information

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IPC IPC(8): C12N15/12C12N15/26C07K14/435C12N15/62A61K38/17A61P35/00
Inventor 吴文芳倪剑锋杨立泉吕安国
Owner SHENYANG INST OF APPLIED ECOLOGY - CHINESE ACAD OF SCI
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