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De-fucosylated fully humanized monoclonal antibody and applications thereof

A monoclonal antibody, fucosylation technology, applied in applications, antibodies, antiviral agents, etc., can solve the problems of threats, no specific prevention and treatment drugs for MERS-CoV, etc.

Inactive Publication Date: 2017-03-08
FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The outbreak of MERS poses a serious threat to global public health. So far, there is no vaccine or specific preventive drug against MERS-CoV

Method used

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  • De-fucosylated fully humanized monoclonal antibody and applications thereof
  • De-fucosylated fully humanized monoclonal antibody and applications thereof
  • De-fucosylated fully humanized monoclonal antibody and applications thereof

Examples

Experimental program
Comparison scheme
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Embodiment 1

[0045] Example 1 Preparation, expression, purification and detection of antibodies

[0046] The preparation, expression, purification of Fab and IgG1 antibody, and the method of converting Fab into IgG1 were carried out according to the method reported in the literature (3);

[0047] The m336-gH gene line was obtained by mutating the 59th amino acid of the heavy chain variable region of m336 from S to N through a site-directed mutagenesis kit, and was prepared using expression and purification conditions similar to those of m336;

[0048] The specific method of Fab preparation is as follows: transfer the plasmid containing the clone m336 or m336-gH into HB2151 competent cells, pick a single colony from the ampicillin plate grown overnight, inoculate the SB bacterial culture medium, and express under the induction condition of 30 degrees IPTG 10-12 hours; Bacteria are harvested and Fabs are purified from them. IgG1 antibody was prepared and expressed by stably transfected and ...

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Abstract

The present invention belongs to the technical field of biology, and relates to a de-fucosylated fully humanized monoclonal antibody for Middle East Respiratory Syndrome Coronavirus (MERS-CoV), an antigen binding fragment, and applications thereof, wherein the de-fucosylated fully humanized monoclonal antibody is mainly characterized in that the de-fucosylated fully humanized monoclonal antibody has an enhanced antibody-dependent cell-mediated cytotoxicity (ADCC) effect, wherein the effect comprises cleaving the cells expressing MERS-CoV S protein. The present invention further relates to a method for making cells express the de-fucosylated MERS-CoV fully humanized monoclonal antibody by inhibiting fucosyltransferase activity. According to the present invention, by using the antibody, the Middle Eastern respiratory syndrome can be clinically prevented or treated.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to defucosylated fully human monoclonal antibodies, in particular to defucosylated fully human monoclonal antibodies against Middle East Respiratory Syndrome Coronavirus (MERS-CoV), Antigen-binding fragments thereof, and uses thereof. Background technique [0002] The prior art discloses that Middle East Respiratory Syndrome Coronavirus (Middle East Respiratory SyndromeCoronavirus, MERS-CoV) is a novel coronavirus that was discovered in June 2012 by Saudi Arabian virologist Ali Moh Zaki in a patient who died of acute pneumonia and Airway epithelial cells isolated from a 60-year-old male patient with acute renal failure. The above MERS patients showed clinical symptoms very similar to SARS, so they were called "SARS-like virus" in the early stage. The main symptoms were fever, cough, shortness of breath, and a high rate of acute renal failure , the fatality rate is currently more than 30...

Claims

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Application Information

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IPC IPC(8): C07K16/10C12N15/13G01N33/68G01N33/577G01N33/569A61K39/42A61P31/14
Inventor 应天雷D.S.迪米特洛夫姜世勃
Owner FUDAN UNIV
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