Method for preparing immunoglobulin G by rapidly extracting plasma of COVID-19 patient in rehabilitation period

A COVID-19, immunoglobulin technology, applied in the direction of serum immunoglobulin, immunoglobulin, antiviral immunoglobulin, etc., can solve the problem of low antibody titer

Active Publication Date: 2020-12-01
深圳博雅感知药业有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, it has been found that even when plasma is separated in a conventional manner using a closed multicellul

Method used

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  • Method for preparing immunoglobulin G by rapidly extracting plasma of COVID-19 patient in rehabilitation period
  • Method for preparing immunoglobulin G by rapidly extracting plasma of COVID-19 patient in rehabilitation period
  • Method for preparing immunoglobulin G by rapidly extracting plasma of COVID-19 patient in rehabilitation period

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Example 1, rapid separation of plasma from convalescent patients with COVID-19

[0054] 1. Peripheral blood (more than 250mL) donated by a COVID-19 patient (in this paper, the patient is marked as patient U) collected by venipuncture during the convalescence period is placed in a blood collection bag (the collection bag is filled with anticoagulant citrate Natrate), put the blood collection bag on a horizontal shaker and mix well for 15 minutes; Recovered persons after confirmation of antibody titer / serum neutralizing antibody titer]

[0055] 2. Using ThermoGenesis Corp.'s closed automatic multi-cell component separation system, the Multicomponent Automated Cell Separation System (MACSS), insert the plastic needle of the disposable separation cup into the sterile interface on the blood collection bag, and hang the blood collection bag 60ml of blood naturally flows into the central compartment of the separation cup; 0.6ml of separation aid is pre-added in the central c...

Embodiment 2

[0066] Embodiment 2, the separation process of preparing component I+II+III and component II and component I+III from raw plasma,

[0067] 1. Preparation of buffer solution:

[0068] ①Ph4.75 0.15mol / L phosphate buffer: Na required per 1kg 2 HPO 4 •12H 2 O is 53g, glacial acetic acid 27mL adjusts the pH to 4.75, and the obtained buffer is referred to as Buffer A;

[0069] ②Ph5.25 0.5mol / L phosphate buffer: Na required per 1kg 2 HPO 4 •12H 2 O is 160.6g, 24mL of glacial acetic acid is used to adjust the pH to 5.25, and the obtained buffer is referred to as Buffer B;

[0070] ③ Component I+III balance liquid for press filtration: 0.13kg of 95% ethanol, Na 2 HPO 4 •12H 2 O is 1.9g, 0.39mL of glacial acetic acid;

[0071] ④1mol / L sodium chloride solution: 58.5g of NaCl is required for every 1kg, add water to 1kg;

[0072] ⑤1mol / L sodium bicarbonate solution: 84.0g of NaCl is required for every 1kg, add water to 1kg;

[0073] ⑥1mol / L HCL solution: make 10.0L 1.0mol / L H...

Embodiment 3

[0084] Example 3, Precipitation and Dissolution of Component II and Purification of Immunoglobulin G by Column Chromatography

[0085] 1. Preparation of buffer solution:

[0086] ① Chromatographic equilibrium solution: add Na per 1kg 2 HPO 4 •12H 2 O 3g, 0.27mL of glacial acetic acid, add low-temperature water for injection to dissolve and adjust the pH to 6.6-7.0, conductivity 1.3-1.5ms / cm, temperature 0-0.5°C;

[0087] ②Ph4.0 phosphate buffer: add Na per 1kg 2 HPO 4 •12H 2 O 17.9g, NaCl 29.2g, glacial acetic acid about 1.5mL, dissolved in low temperature water for injection;

[0088] ③ pH4.0 acetic acid eluent: add NaAc•3H per 1kg 2 O 20.7g, NaCl 58.5g, glacial acetic acid 20.5mL, add water for injection to adjust pH=4.0;

[0089] 2. Dissolve and dilute the component II precipitate obtained in Example 2 with 10 times of water for injection, the dissolution temperature is 0-4°C, the dissolution time is ≥2h, and after dissolution, adjust the pH to 6.6-7.0, the conduc...

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Abstract

The invention relates to a method for preparing immunoglobulin G by rapidly extracting plasma of a COVID-19 patient in a rehabilitation period, and more particularly, to a method for preparing intravenous injection COVIDI-19 immunoglobulin G by rapidly separating component plasma from whole blood by using a closed multicellular component automatic separation system (MACSS), for example, to prepareintravenous injection superimmunoglobulin. The intravenous immunoglobulin can be used for treating patients infected by novel coronavirus. The method comprises the following steps: separating human peripheral blood into three component layers, namely an erythrocyte layer, a cell concentration layer and a plasma layer, by using a closed multi-cell component automatic separation system, and then carrying out virus inactivation on the obtained plasma to obtain plasma capable of being used for preparing intravenous immunoglobulin G. The invention also relates to a method for preparing an intravenous immunoglobulin G preparation by using the plasma component obtained by the above method. According to the method, the plasma obtaining speed is high, and the antibody titer of the prepared plasmaand intravenous immunoglobulin G is high.

Description

technical field [0001] The invention belongs to the field of medical biotechnology, and specifically relates to a method for rapidly separating plasma components from whole blood to prepare intravenous COVID-19 immunoglobulin. In particular, it involves the use of a closed multicomponent automated cell separation system, the Multicomponent Automated Cell Separation System (MACSS, ThermoGenesisCorp.), to rapidly separate component plasma from whole blood for the preparation of intravenous COVID-19 immunoglobulin G, for example using To prepare hyperimmune globulin for intravenous injection. The intravenous injection of COVID-19 immune globulin can be used for the treatment of patients infected with the novel coronavirus COVID-19. Background technique [0002] Coronavirus disease 2019 (Coronavirus Disease 2019, COVID-19), also known as novel coronavirus, is a disease caused by severe acute respiratory syndrome coronavirus 2 (Severe Acute Respiratory Syndrome Coronavirus 2, SA...

Claims

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Application Information

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IPC IPC(8): C07K16/10C07K16/06A61K39/42A61P31/14
CPCA61K2039/505A61P31/14C07K16/065C07K16/10C07K2317/76
Inventor 魏卿肖海蓉刘庆喜
Owner 深圳博雅感知药业有限公司
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