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P28-Fc fusion protein and coding nucleic acid molecule, recombinant cell expression carrier, recombinant and application thereof

A p28-fc, fusion protein technology, applied in the direction of peptide/protein components, fusion polypeptide, recombinant DNA technology, etc., can solve the problem of inability to obtain soluble IL-27p28 protein

Inactive Publication Date: 2019-04-19
王仁喜
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] In order to overcome the defect that soluble IL-27p28 protein cannot be obtained in the prior art, the present invention provides a p28-Fc fusion protein and its encoding nucleic acid molecule, recombinant expression vector, and recombinant cell. The present invention combines human IL-27p28 protein with Fc protein Recombination after transformation, the obtained fusion protein can be soluble expressed in insect cells, so as to obtain the fusion protein containing IL-27p28 in the correct configuration

Method used

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  • P28-Fc fusion protein and coding nucleic acid molecule, recombinant cell expression carrier, recombinant and application thereof
  • P28-Fc fusion protein and coding nucleic acid molecule, recombinant cell expression carrier, recombinant and application thereof
  • P28-Fc fusion protein and coding nucleic acid molecule, recombinant cell expression carrier, recombinant and application thereof

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Embodiment 1

[0069] (1): Construction of hp28-hIgG4Fc insect expression vector

[0070] Expression vector construction process:

[0071] 1. Query the base sequence and amino acid sequence of human IL-27A, that is, the IL-27p28 subunit containing the signal peptide, through the Pubmed database

[0072] Human IL-27p28 cDNA (SEQ ID NO.7)

[0073]

[0074]

[0075] Wherein, the part in bold is the coding base of the signal peptide of human IL-27p28.

[0076] Human IL-27p28 amino acid sequence (SEQ ID NO.8)

[0077] MGQTAGDLGWRLSLLLLPLLLVQAGVWGFPRPPGRPQLSLQELRREFTVSLHLARKLLSEVRGQAHRFAESHLPGVNLYLLPLGEQLPDVSLTFQAWRRLSDPERLCFISTTLQPFHALLGGLGTQGRWTNMERMQLWAMRLDLRDLQRHLRFQVLAAGFNLPEEEEEEEEEEEEERKGLLPGALGSALQGPAQVSWPQLLSTYRLLHSLELVLSRAVRELLLLSKAGHSVWPLGFPTLSPQP

[0078] Wherein, the bold part is the amino acid sequence of the signal peptide of human IL-27p28.

[0079] 2. Analyze the codon preference of IL-27A by computer

[0080] Analysis results such as figure 1 As shown, the signal pept...

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Abstract

The invention provides a P28-Fc fusion protein and a coding nucleic acid molecule, a recombinant expression carrier, a recombinant cell and application thereof, and relates to the technical field of drugs. The p28-Fc fusion protein is composed of hIL-27p28 and Fc. There exists base sequence which is adverse to mammal expression in a human IL-27p28 signal peptide coding area, in order to obtain theIL-27p28 with a correct configuration, a Hongeybee melittin secretion signal peptide is included in hIL-27p28 to replace the human IL-27p28 signal peptide, and therefore the problem that the solubleIL-28p28 with the correct configuration cannot be obtained in heterologous expression is prevented. The invention further provides the application in preparing drugs for treating autoimmune diseases of the p28-Fc fusion protein and the coding nucleic acid molecule, recombinant expression carrier or recombinant cell thereof.

Description

technical field [0001] The invention relates to the field of pharmaceutical technology, in particular to p28-Fc fusion protein and its encoding nucleic acid molecule, recombinant expression vector, recombinant cell and application. Background technique [0002] Interleukin-27 (IL-27) is a member of the IL-12 family (IL-12, IL-23, IL-27 and IL-35). IL-12 family cytokines are composed of an α chain (p19, p28 or p35) and a β chain (p40 or EBI3), for example IL-27 is composed of p28 and EBI3 (Epstein-Barr virus-induced gene 3) [Pflanz, S. et al. (2002) Immunity 16:779.]. IL-27p28 is also known as IL27, p28, IL30, IL-27, IL27p28, etc. [0003] p28 is a polypeptide similar to p35 of IL-12, and EBI3 is a protein similar to p40 of IL-12 / IL-23. IL-27 binds to the heterodimeric receptor complex formed by TCCR / WSX-1 and gp130, a receptor subunit common to cytokines of the IL-6 family [Pflanz, S. et al. (2004 ) J. Immunol. 172:2225]. [0004] IL-27 is a major suppressive cytokine b...

Claims

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Application Information

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IPC IPC(8): C07K19/00C12N15/62C12N15/85C12N5/10A61K38/20A61P37/02A61P25/00A61P17/06A61P27/02
CPCA61K38/00A61P17/06A61P25/00A61P27/02A61P37/02C07K14/5434C07K2319/30C12N2510/02
Inventor 王仁喜
Owner 王仁喜