Novel erythrocyte membrane monoclonal antibody production method and processing device thereof

A technology of red blood cell membrane and production method, applied in the field of biomedicine, can solve the problems of low specificity, increased production cost, poor control accuracy of production method, etc., achieve high cancer cell uptake rate, avoid bubble generation, and sample loading effect better results

Active Publication Date: 2020-07-31
江苏帆博生物制品有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In this method, the control accuracy of the process production method is poor, the speed and time of the overall temperature, stirring, centrifugation, and peristalsis are difficult to control, and the production efficiency is low.
[0008] In summary, the problems existing in the prior art are: (1) The existing erythrocyte membrane monoclonal antibody is not highly sensitive and specific, is not easy to be judged, and cannot be mass-produced
(2) PEGylation has become the gold standard for increasing circulation time, but there are still some problems, such as insufficient product purity, high selectivity for modified molecules, possible increase in drug toxicity, and difficulty in excreting large molecular PEGs from the body, etc.
[0009] Difficulty in solving the above technical problems: The difficulty in solving the above problems lies in the fact that erythrocyte membrane monoclonal antibodies cannot be mass-produced, which leads to an increase in production costs and limits its further application

Method used

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  • Novel erythrocyte membrane monoclonal antibody production method and processing device thereof
  • Novel erythrocyte membrane monoclonal antibody production method and processing device thereof
  • Novel erythrocyte membrane monoclonal antibody production method and processing device thereof

Examples

Experimental program
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Effect test

preparation example Construction

[0075] In S109 provided by the present invention, the preparation method of the antibody nanoparticles comprises the following steps:

[0076] 1) Preparation of erythrocyte membrane: after fully anesthetizing SD rats, take blood from the heart, transfer the blood to an EP tube containing sodium heparin; centrifuge the whole blood, discard the supernatant, leave a dark red precipitate, wash with PBS, and centrifuge , and then washed several times; discard the supernatant, leave the precipitate, take PBS and blow it away, take 0.2mM EDTA and put it into a centrifuge tube to mix well, after fully hemolysis, add 20*PBS, centrifuge, and centrifuge for hemolysis several times; discard The supernatant, the precipitate is the red blood cell membrane.

[0077] 2) Preparation of antibody nanoparticles by isoelectric point precipitation: put the antibody into a dialysis bag, dialyze in ultrapure water, collect the antibody in the dialysis bag, freeze-dry, and store at -20°C; take the fre...

Embodiment 1

[0126] Example 1: Production method of novel erythrocyte membrane monoclonal antibody

[0127] 1. Workflow:

[0128] 1.1 Preparation:

[0129] 1.1.1 Reagent preparation:

[0130] Equilibration buffer: 10 mM Tris-NaCl (pH8.3).

[0131] Dissociation buffer: 0.1M citric acid (pH 4.0).

[0132] Regeneration buffer: 0.1M glycine (pH2.7).

[0133] Dialysis buffer: 20 mM Tris-NaCl (pH7.5) or 10 mM PBS (pH7.4).

[0134] pH neutralizing solution: 2M Tris (pH8.0).

[0135] 1.1.2 Equipment preparation: Protein A column, low-temperature high-speed centrifuge, balance, peristaltic pump.

[0136] 1.2 Operation steps:

[0137] 1.2.1 Sample preparation: The frozen ascites was melted at 10-15°C the day before, and filtered with absorbent cotton soaked in triple-distilled water to remove oil, and the filtered ascites was centrifuged at 4°C at 10,000 rpm for 30 minutes to collect the supernatant.

[0138]Column preparation: Fill with an appropriate amount of Protein A filler, wash with p...

Embodiment 2

[0169] Embodiment 2: test report, as follows:

[0170]

[0171]

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Abstract

The invention belongs to the technical field of biomedicine, and discloses a novel erythrocyte membrane monoclonal antibody production method. The method comprises the following steps: dissolving andfiltering ascites, adding dextran sulfate and calcium chloride, performing centrifuging, leaching, balancing, diluting, cleaning, dissociating, dialyzing and incubating, preparing antibody nanoparticles, wrapping the nanoparticles with erythrocyte membranes, and performing filtering and storing. The production method is simple to operate, the process is easy to control, and the yield is high; a novel erythrocyte membrane monoclonal antibody produced by the method is high in sensitivity and specificity, and the result is easy to judge; the monoclonal antibody also has the characteristics of high stability, high cancer cell uptake rate, low reticuloendothelial system clearance rate and obvious prolonging of in-vivo circulation action time; and a processing device can accurately control the production method on the basis of the preset process, improves the production efficiency, reduces the complexity of production operation, improves the economic benefit, can achieve intelligent control,and reduces the labor investment.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a method for producing a novel erythrocyte membrane monoclonal antibody and a processing device thereof. Background technique [0002] At present, the existing technology commonly used in the industry is as follows: Compared with traditional small molecule anticancer drugs, antibodies are increasingly becoming one of the most important anticancer drugs due to their high specificity and strong affinity. In contrast, although small molecule anticancer drugs have certain efficacy and are still widely used in chemotherapy, they still have disadvantages such as non-specific toxicity (due to targeting all differentiated cells) and small therapeutic window. In addition, the drug resistance of small molecule anticancer drugs is also a common problem in their clinical use. These shortcomings have brought a lot of pain to chemotherapy patients. Antibodies have attracted mo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/00C07K16/40C07K1/36C07K1/30C07K1/16
CPCC07K1/16C07K1/30C07K1/36C07K16/00C07K16/40C07K2317/35C07K2317/56
Inventor 张杰李永刚
Owner 江苏帆博生物制品有限公司
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