Optimized Fc fragment and optimizing method and application thereof

An optimization method and fragment technology, applied in chemical instruments and methods, biochemical equipment and methods, applications, etc., can solve the problems of poor thermal stability and anti-aggregation ability of Fc fragments, and achieve improved stability and anti-aggregation, Good anti-aggregation ability and good stability

Inactive Publication Date: 2015-11-11
WUHAN INST OF VIROLOGY CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Solve the technical problems of poor thermal stability and anti-aggregation ability of existing Fc fragments

Method used

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  • Optimized Fc fragment and optimizing method and application thereof
  • Optimized Fc fragment and optimizing method and application thereof
  • Optimized Fc fragment and optimizing method and application thereof

Examples

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Embodiment 1

[0036] The optimized Fc fragment of the present invention uses the Fc gene fragment as the backbone, modifies it, cuts off the 7 irregularly coiled amino acids at the N-terminal of the Fc fragment, and finally obtains an optimized Fc fragment with improved stability and anti-aggregation performance. Fc clone, named Fcs. The optimized Fc fragment of the present invention is represented by "Fcs" below. Wherein, the Fc gene fragments may be Fc fragments in immunoglobulins of different species.

[0037] Taking the Fc fragment of human immunoglobulin as the object of transformation, the optimization method of the optimized Fc fragment of the present invention is described, including the following steps:

[0038] In order to facilitate the comparison of various properties of the Fcs (optimized Fc fragment) of the present invention and Fc, the steps of Fc amplification and expression purification are also described in the optimization method of Example 1.

[0039] Step 1. Amplify F...

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Abstract

The invention provides an optimized Fc fragment and an optimizing method and application thereof. The optimized Fc fragment is obtained by removing seven irregular curved amino acids at N ends of an Fc fragment. The optimizing method comprises the amplifying steps that RT-PCR is utilized for amplifying wild type Fc out of a blood sample, a primer is designed to cut off the seven amino acids at the N ends of the wild type Fc, and the optimized Fc fragment is amplified out, and further comprises expressing and purifying steps. The optimized Fc fragment and the optimizing method and application thereof are used for applying improvement with the Fc fragment. A provided Fcs framework is better in stability, and the producing, purifying and storing cost of a monoclonal antibody or Fc fusion protein can be lowered. The gathering resisting capacity is better, and clinical use risks caused by protein gathering can be lowered. The optimized Fc fragment framework can be applied to complete IgG or relevant Fc fusion protein and used for further studying the influence on the plasma half-life of relevant biological agents.

Description

technical field [0001] The present invention relates to the technical field of biomedical engineering, in particular to an optimized Fc fragment and its optimization method and application. Background technique [0002] With its strong specificity and high sensitivity, antibodies are widely used in various fields, especially in life science research and clinical treatment such as ELISA, Western blot, immunofluorescence analysis, rapid diagnosis of diseases, and as biological agents to treat various diseases. Therefore, since entering the 21st century, people have begun to pay more and more attention to the research and development and clinical application of therapeutic antibody drugs, because it has less toxic and side effects on the human body, has natural and highly specific curative effects, and has created a huge social benefits and economic benefits. [0003] At the same time, fusion proteins based on Fc fragments not only have good tissue permeability due to their re...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/00C12N15/13C12N15/10C12N15/70C07K19/00
Inventor 龚睿陈小波张哲
Owner WUHAN INST OF VIROLOGY CHINESE ACADEMY OF SCI
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