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Human platelet collagen receptor resistant human-derived monoclonal antibody fragment and application thereof

A monoclonal antibody, collagen technology, applied in the direction of anti-animal/human immunoglobulin, anti-receptor/cell surface antigen/cell surface determinant immunoglobulin, antibody, etc., can solve the problem of high preparation cost and complex technology and other problems, to achieve the effect of reducing the preparation cost and shortening the preparation time

Inactive Publication Date: 2013-11-20
ANHUI NEW MARK SCI TECH
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  • Description
  • Claims
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Problems solved by technology

The technology required for the whole process is very complex and the preparation cost is high

Method used

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  • Human platelet collagen receptor resistant human-derived monoclonal antibody fragment and application thereof
  • Human platelet collagen receptor resistant human-derived monoclonal antibody fragment and application thereof

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Embodiment Construction

[0035] 1. Use normal human platelets as the antigen source (immunogen) of the collagen receptor "glycoprotein-6", and patients with thrombocytopenic purpura (lack of platelet collagen receptor "glycoprotein-6") as the immune receptor , prepare B lymphocytes expressing anti-GPVI antibody from the patient, and extract nucleotides from it:

[0036] (1) Screening patients: In cooperation with the hematology department of the hospital, 15 ml of blood was drawn with sodium citrate as an anticoagulant for patients with thrombocytopenic purpura after obtaining the consent of the patients themselves and their family members, and obtained by low-speed centrifugation. Supernatant (platelet-rich plasma, specific methods can refer to Sun, B et al..J..Cardiovascular Pharmacol..40:557--585, 2002). Take 400 microliters of the resulting supernatant and put it on a platelet aggregation instrument to preheat at 37 degrees Celsius, and then use 10 or 20 micromolar (μM) adenosine diphosphate (ADP)...

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Abstract

The invention relates to the technical field of biology, also relates to the bio-pharmaceutical technology, and particularly relates to a human platelet collagen receptor ''Glycoprotein VI'' (or GPVI) resistant human-derived monoclonal antibody fragment and application thereof in medicines. The monoclonal antibody fragment is human derived, and comprises polypeptide amino acid sequences comprising three heavy chain variable region HV sequences and three light chain variable region LV sequences, antibody fragment scFv(single chain fragment variable) or other polypeptides recombined by the monoclonal antibody fragment can be used as a blocking agent of collagen receptor ''Glycoprotein VI'' for application in preparation of medicines for treatment or prevention of thrombotic cardio cerebrovascular diseases, the monoclonal antibody fragment can be used in the preparation of antithrombotic medicines resisting collagen to activate platelet, or in the preparation of detection reagents and the like for detecting whether free ''Glycoprotein VI'' exists. In the preparation of the medicines, a very complex humanized reconstruction process is omitted.

Description

technical field [0001] The present invention relates to the field of biotechnology, and also relates to biopharmaceutical technology. It specifically relates to a humanized monoclonal antibody fragment against human platelet collagen receptor "glycoprotein-6" (Glycoprotein VI or GPVI) and its application in medicine. Background technique [0002] The adhesion, activation and aggregation of platelets at damaged blood vessels will eventually lead to thrombus formation and bleeding stop, which can avoid excessive blood loss. This is one of the physiological protection mechanisms of normal people. However, in the case of vascular disease, the thrombus formed by platelet activation will block the blood vessel, causing ischemia or even necrosis of downstream tissues, and myocardial infarction or stroke. Therefore, antithrombotic therapy is widely used in medicine. For example, it is used for the prevention of thrombosis caused by vascular endothelial injury, such as medication ...

Claims

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Application Information

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IPC IPC(8): C07K16/28A61K39/395A61P7/02A61P9/10G01N33/68G01N33/577
Inventor 孙冰张协平徐镜
Owner ANHUI NEW MARK SCI TECH
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