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Screening method and application of bacillus subtilis proprotease 9 targeted binding protein

A Bacillus subtilis and targeted combination technology, applied in the field of biomedicine, can solve the problems of insufficient clinical promotion of drugs, non-targeted immune response, complex preparation process, etc., and achieve the effects of avoiding repeated freezing and thawing, easy storage, and simple process

Pending Publication Date: 2022-05-06
刘博巽
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Problems solved by technology

However, there are certain inherent defects in mAb therapy, including: mAb contains Fc segment, whether it is humanized or not, it may have cross-linking interaction with immune cells, and there may even be an unintended immune response; mAb can induce drug resistance in vivo Antibody (ADA) is possible (bococizumab and evolocizumab have been reported); and the ADA of bococizumab can produce cross-resistance with other similar drugs, which has become one of the important reasons for the failure of the market; mAb preparation depends on eukaryotic cells, preparation The complex process and high cost lead to insufficient clinical promotion of such drugs, which has led to doubts in the academic circles about the efficacy / expenditure ratio of mAbs

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  • Screening method and application of bacillus subtilis proprotease 9 targeted binding protein
  • Screening method and application of bacillus subtilis proprotease 9 targeted binding protein
  • Screening method and application of bacillus subtilis proprotease 9 targeted binding protein

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Embodiment Construction

[0012] The present invention will be described in detail below in conjunction with the accompanying drawings and specific embodiments.

[0013] like Figures 1 to 4 Shown, the operation process of the present invention is as follows:

[0014] 1. Screening of PCSK9 targeting binding proteins in phage display library:

[0015] (1) Add 2ml of 100ug / ml recombinant human PCSK9 protein solution into a small dish that has been placed in the opening and irradiated with ultraviolet rays for 1 hour to coat overnight; after coating, the liquid in the dish is fully sucked out, and the liquid containing 0.5% bovine serum albumin is used. (BSA) in 0.5% Tween in phosphate buffered saline (0.5% PBST) at 37°C for 1 h.

[0016] (2) After repeated washing with 2ml of 0.5% PBST for several times, 2ml of phage display peptide library was added to the small dish, and incubated at room temperature for 2h to complete the incubation process.

[0017] (3) Carefully suck out the phage solution in the...

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Abstract

The invention discloses preparation and application of a bacillus subtilis proprotease 9 (PCSK9) targeted binding protein. The PCSK9 targeted binding protein is screened and prepared through a phage display peptide technology, and a convenient, efficient, stable and low-cost choice is provided for treatment and management of hypercholesteremia and atherosclerotic cardiovascular diseases.

Description

technical field [0001] The invention belongs to the technical field of biomedicine and relates to the preparation of a bacillus subtilis proproteinase 9 (PCSK9) targeting binding protein and its application in hypercholesterolemia and atherosclerotic cardiovascular disease (ASCVD). Background technique [0002] Hypercholesterolemia is an important pathophysiological basis of atherosclerotic diseases. High-dose statins are recommended in lipid-lowering guidelines as a blood cholesterol management method for high-risk patients (such as ASCVD), but there are contradictions between treatment goals and discomfort reactions, including: (1) Guidelines require ASCVD patients with serum LDL-cholesterol < 1.8 mmol / L, this target is extremely strict for patients with high baseline LDL-cholesterol levels and monotherapy; (2) Large doses of statins often have the risk of adverse reactions such as liver damage and striated muscle damage, Some patients are difficult to tolerate; (3) Mo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10C12N15/70C07K14/00A61K38/16A61P9/10A61P3/06C12R1/19
CPCC12N15/1037C12N15/70C07K14/00A61P9/10A61P3/06A61K38/00
Inventor 刘博巽
Owner 刘博巽
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