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Preparation method and application of monoclonal antibody

A monoclonal antibody, a single technology, applied in the direction of antibodies, antiviral agents, antiviral immunoglobulins, etc., can solve the problems of high cost, low concentration of recombinant antibodies, unsuitable for large-scale production and application, and achieve cost reduction and recombination Effect of Elevated Antibody Concentration

Inactive Publication Date: 2015-08-12
上海泰坦科技股份有限公司
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Problems solved by technology

[0010] Although the above-mentioned prior art discloses some preparation methods of fully humanized antibodies against infectious disease pathogens, which can meet certain needs, there are still certain defects in these methods: the obtained recombinant antibodies have low concentration and high cost, and are not suitable for industrial production. mass production applications

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Embodiment Construction

[0025] The present invention will be described in detail below in conjunction with specific embodiments. The following examples will help those skilled in the art to further understand the present invention, but do not limit the present invention in any form. It should be noted that those skilled in the art can make several modifications and improvements without departing from the concept of the present invention. These all belong to the protection scope of the present invention.

[0026] The following examples relate to the preparation of human monoclonal antibodies to hepatitis C virus E1 protein, which consists of the following steps:

[0027] Step 1. The hepatitis C virus E1 protein is the main protective antigen of the virus. Neutralizing monoclonal antibodies against the E1 protein can be used for the treatment of patients with hepatitis C and liver cancer. Screening and cloning of the neutralizing monoclonal antibodies against the hepatitis C virus E1 has Significant ...

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Abstract

The invention relates to a preparation method and an application of a monoclonal antibody. The preparation method comprises the following steps: 1, obtaining a B cell, enriching, and purifying; 2, binding a labeled antigen to a specific B cell; 3, obtaining an antigen specific B cell; 4, amplifying the variable region gene of an antibody; 5, constructing an expression vector; and 6, expressing a recombinant antibody and purifying: transfecting a 293T cell in the logarithmic phase by using purified expression vector DNA, centrifuging to remove cell fragments, and carrying out protein G column purification on the obtained centrifuge solution to obtain a recombinant monoclonal antibody solution. The preparation method substantially rises the concentration of the recombinant monoclonal antibody, reduces the cost, and is suitable for industrial large-scale production application. The obtained monoclonal antibody can specifically neutralize hepatitis C E1 antigens, and can be used for preparing drugs for preventing and treating hepatitis C.

Description

technical field [0001] The invention belongs to the technical field of monoclonal antibodies, and in particular relates to a preparation method and application of monoclonal antibodies. Background technique [0002] The main function of antibodies is to combine with antigens (including foreign and self), so as to effectively remove foreign substances such as microorganisms and parasites invading the body, especially monoclonal antibodies, because of their high uniformity and specific antigen expression. Therefore, it is widely used in the fields of immunological analysis, radioimmunoimaging and immune-guided therapy. [0003] The development of monoclonal antibodies has gone through four stages, namely: murine monoclonal antibodies, chimeric monoclonal antibodies, humanized monoclonal antibodies and fully human monoclonal antibodies. [0004] Mouse-derived monoclonal antibody: mouse hybridoma monoclonal antibody mainly fuses B cells from immunized mice with myeloma cells, a...

Claims

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Application Information

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IPC IPC(8): C12N15/85C07K16/10A61K39/395A61P31/14
Inventor 谢应波张庆张华徐肖冰罗桂云
Owner 上海泰坦科技股份有限公司
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