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Lyophilized tachypleus amebocyte lyophilized microspheres as well as preparation method and application thereof

A Limulus reagent and microsphere technology, applied in the field of Limulus reagent freeze-dried microspheres and preparation thereof, can solve the problems of cuts, difficult to accurately divide or simplify operations, difficult to integrate instant inspection, etc. Low deviation, the effect of improving hardness

Pending Publication Date: 2021-07-09
DYNAMIKER BIOTECH TIANJIN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] It can be seen from the above that the conventional vacuum freeze-drying method freeze-dries the LAL reagent in a vial or ampoule to obtain a powdered LAL reagent. Point-of-care testing (POCT) products such as test cards or chips. In addition, the seal of the container needs to be broken during operation, and there are risks such as cuts

Method used

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  • Lyophilized tachypleus amebocyte lyophilized microspheres as well as preparation method and application thereof
  • Lyophilized tachypleus amebocyte lyophilized microspheres as well as preparation method and application thereof
  • Lyophilized tachypleus amebocyte lyophilized microspheres as well as preparation method and application thereof

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Effect test

Embodiment 1

[0049] This embodiment provides a kind of limulus reagent freeze-dried microsphere, the preparation method of described limulus reagent freeze-dry microsphere comprises the following steps:

[0050] (1) Take the raw material of Limulus amoeba cells out of the freezer stored at low temperature (-20°C), and place it at room temperature to melt slowly, so as to prevent the infusion bottle containing the raw material from breaking due to too fast melting;

[0051] (2) Perform high-speed homogenization and crushing of the completely melted Limulus amoeba cell raw material, centrifuge to collect the supernatant in a low-temperature high-speed centrifuge at 4°C and 8000rpm, discard cell debris and other substances, and add three Chloromethane (chromatographically pure) was extracted, magnetically stirred for 20 min, and the supernatant was collected by centrifugation in a low-temperature high-speed centrifuge at 4°C and 12,000 rpm to obtain a Limulus amoeba cell extract;

[0052] (3)...

Embodiment 2

[0059] This embodiment provides a kind of limulus reagent freeze-dried microsphere, the preparation method of described limulus reagent freeze-dry microsphere comprises the following steps:

[0060] (1) Take out the raw material of Limulus amoeba cells from the freezer stored at low temperature, place it at room temperature and slowly melt it, so as to prevent the infusion bottle containing the raw material from breaking due to too fast melting speed;

[0061] (2) Perform high-speed homogenization and crushing of the completely melted Limulus amoeba cell raw material, centrifuge to collect the supernatant in a low-temperature high-speed centrifuge at 4°C and 8000rpm, discard cell debris and other substances, and add three Chloromethane (chromatographically pure) was extracted, magnetically stirred for 20 min, and the supernatant was collected by centrifugation in a low-temperature high-speed centrifuge at 4°C and 12,000 rpm to obtain a Limulus amoeba cell extract;

[0062] (3)...

Embodiment 3

[0069] This embodiment provides a kind of limulus reagent freeze-dried microsphere, the preparation method of described limulus reagent freeze-dry microsphere comprises the following steps:

[0070] (1) Take out the raw material of Limulus amoeba cells from the freezer stored at low temperature, place it at room temperature and slowly melt it, so as to prevent the infusion bottle containing the raw material from breaking due to too fast melting speed;

[0071] (2) Perform high-speed homogenization and crushing of the completely melted Limulus amoeba cell raw material, centrifuge to collect the supernatant in a low-temperature high-speed centrifuge at 4°C and 8000rpm, discard cell debris and other substances, and add three Chloromethane (chromatographically pure) was extracted, magnetically stirred for 20 min, and the supernatant was collected by centrifugation in a low-temperature high-speed centrifuge at 4°C and 12,000 rpm to obtain a Limulus amoeba cell extract;

[0072] (3)...

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Abstract

The invention relates to lyophilized tachypleus amebocyte lysate microspheres as well as a preparation method and application thereof. The lyophilized tachypleus amebocyte lyophilized microspheres comprise amebic cell extracts of tachypleus amebocyte lyophilized microspheres and an active agent, wherein the active agent comprises polyvinylpyrrolidone, lactose, mannitol, raffinose, polyethylene glycol and glycine. All the components cooperate with one another to promote the formation of the tachypleus amebocyte lysate, improve hardness of the formed tachypleus amebocyte lysate and protect activity of the tachypleus amebocyte lysate in the precooling period, the freeze-drying period and the storage period, so the tachypleus amebocyte lysate freeze-dried microspheres are high in hardness, smooth in surface, not prone to breaking and chipping, high in stability and convenient for subsequent automatic or manual sub-packaging and storage.

Description

technical field [0001] The invention belongs to the technical field of freezing biological products, and relates to a limulus reagent freeze-dried microsphere and a preparation method and application thereof. Background technique [0002] Limulus reagent is a biological monitoring reagent made from the extract of blood amoeba cells of the marine organism Limulus limulus. Limulus reagent contains factor C, factor B and factor G that can be activated by bacterial endotoxin (lipopolysaccharide) and fungal glucan , coagulase and coagulogen, etc., can accurately and quickly detect whether the sample contains bacterial endotoxin and (1,3)-β-D-glucan qualitatively or quantitatively. [0003] At present, Limulus reagent is widely used in the fields of pharmacy, clinical and scientific research, for the detection of bacterial endotoxin and fungal glucan, but the Limulus reagent is unstable and easy to contaminate in the liquid state. Currently, the Limulus reagent on the market uses ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/579G01N33/543
CPCG01N33/579G01N33/543Y02A50/30
Inventor 王保学钱芃橙苗苗刘凯刘浩宇
Owner DYNAMIKER BIOTECH TIANJIN
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