Culture medium processing system and method with deproteinization in a filtration container

Patent metadata

Field	Value
Publication number	US-11454640-B2
Application number	US-202016821205-A
Country	US
Kind code	B2
Filing date	Mar 17, 2020
Priority date	Apr 18, 2019
Publication date	Sep 27, 2022
Grant date	Sep 27, 2022

Abstract

The culture medium processing system includes a controller (100) configured to control operation of a sample dispensing part (20), a reagent dispensing part (26) and a transport arm (24) to deproteinize a sample, wherein the controller (100) is configured to dispense a methanol solution into an empty filtration container (50) to perform a conditioning of a filtration filter (52) disposed in the filtration container (50), then dispense a sample into the filtration container (50), add an acetonitrile solution as a deproteinization agent to the sample in the filtration container (50), and then perform a filtration process in the filtration part (30).

First claim

What is claimed is:

1. A culture medium processing system comprising:
an extraction device including a filtration container and a collection container, wherein the filtration container has an open upper surface, a filtration filter, disposed inside the filtration container, for deproteinizing a culture medium for cell culture, and an extraction port, disposed at a lower end of the filtration container, for extracting a filtrate downward that has passed through the filtration filter, the collection container being for collecting the filtrate extracted from the extraction port of the filtration container by being disposed immediately below the filtration container;
an extraction device setting place at which the extraction device is set in an empty state;
a sample container setting place at which sample containers containing culture medium to be deproteinized as samples are set;
a reagent container setting place in which reagent containers, which include a container containing a methanol solution as a reagent and a container containing an acetonitrile solution as a reagent, are set;
a sample dispensing part configured to suck a sample from a sample container set at the sample container setting place and dispense the sucked sample into the filtration container disposed at a predetermined sample dispensing position;
a reagent dispensing part configured to suck desired reagents from the reagent containers set at the reagent container setting place and to dispense the sucked reagents into the filtration container disposed at a predetermined reagent dispensing position;
a stirring part including a stirring port at which the filtration container is set, the stirring part being configured to stir a liquid inside the filtration container set in the stirring port,
a filtration part including a filtration port at which the extraction device is set with the collection container disposed immediately below the filtration container, the filtration part being configured to collect a filtrate into the collection container by performing a filtration process for a solution in the filtration container of the extraction device set at the filtration port;
a transport arm configured to hold the filtration container and/or the collection container of the extraction device set at the extraction device setting place and to transport the held filtration container and/or the held collection container to the sample dispensing position, the reagent dispensing position, the stirring part, and the filtration part; and
a controller configured to control operation of the sample dispensing part, the reagent dispensing part and the transport arm to deproteinize a sample,
wherein the controller is configured to perform conditioning for the filtration filter, the deproteinization, and then the filtration process in the filtration part, the conditioning is performed by sucking the methanol solution from the container containing the methanol solution and dispensing a first amount of the sucked methanol solution into the empty filtration container so as to wet the filtration filter which has been dry, after the conditioning, the deproteinization is performed by dispensing a second amount of a sample into the filtration container and by sucking the acetonitrile solution from the container containing the acetonitrile solution, dispensing a third amount, which is larger than the first amount and second amount, of the acetonitrile solution as a deproteinization agent into the filtration container and stirring the sample and the acetonitrile solution in the filtration container.

2. The culture medium processing system according to claim 1 , wherein the first amount, the second amount, and the third amount are each fixed amount.

3. The culture medium processing system according to claim 1 , wherein
a standard solution container containing an internal standard solution is set at the reagent container setting place, and
the controller is configured to dispense the internal standard solution into the filtration container before dispensing a sample into the filtration container after performing the conditioning for the filtration filter.

4. The culture medium processing system according to claim 3 , wherein the controller is configured to dispense a fixed amount of the internal standard solution into the filtration container.

5. The culture medium processing system according to claim 1 , further comprising an LC part including a sample injection part and an analysis part, wherein the sample injection part is configured to suck a fixed amount of a filtrate collected in the collection container by the filtration process and to injecting the filtrate into an analysis flow path for a liquid chromatography analysis, and the analysis part is configured to perform a liquid chromatography analysis of the filtrate injected into the analysis flow path by the sample injection part.

6. The culture medium processing system according to claim 5 , wherein the sample injection part is configured to dilute the fixed amount of filtrate sucked from the collection container at a predetermined magnification.

7. A method of processing a culture medium using an extraction device, the extraction device including
a filtration container having an open upper surface, a filtration filter, disposed inside the filtration container, for deproteinizing a culture medium for cell culture, and an extraction port, disposed at a lower end of the filtration container, for extracting a filtrate downward that has passed through the filtration filter, and
a collection container for collecting the filtrate extracted from the extraction port of the filtration container by being disposed immediately below the filtration container,
the method of processing the culture medium comprising, in the sequence set forth:
a conditioning step of dispensing a first amount of a methanol solution into the empty filtration container to wet the filtration filter which has been dry;
a sample dispensing step of dispensing a second amount of a culture medium as a sample into the filtration container;
a deproteinization agent dispensing step of dispensing a third amount, which is larger than the first amount and the second amount, of an acetonitrile solution as a deproteinization agent into the filtration container including the filtration filter conditioned by the methanol solution so as to add the acetonitrile solution to the sample in the filtration container;
a stirring step of stirring a liquid in the filtration container after the deproteinization agent dispensing step; and
a filtration step of filtering a solution in the filtration container with the collection container disposed immediately below the filtration container to collect the filtrate in the collection container.

8. The method of processing the culture medium according to claim 7 , wherein the first amount, the second amount, and the third amount are each fixed amounts.

9. The method of processing the culture medium according to claim 7 , further comprising a standard solution dispensing step of dispensing an internal standard solution into the filtration container before the sample dispensing step after the conditioning step.

10. The method of processing the culture medium according to claim 9 , wherein the standard solution dispensing step includes dispensing a fixed amount of the internal standard solution into the filtration container.

11. The method of processing the culture medium according to claim 7 , further comprising:
an injection step of sucking a fixed amount of a filtrate collected in the collection container in the filtration step after the filtration step, and then injecting the sucked filtrate into an analysis flo

Assignees

• Shimadzu Corp

Inventors

• Toyoda Kenichi
• Suzuki Takashi

Classifications

• G01N30/06

  Preparation · CPC title

• G01N30/16

  Injection (G01N30/24 takes precedence) · CPC title

• G01N35/025

  having a carousel or turntable for reaction cells or cuvettes · CPC title

• G01N2030/027

  Liquid chromatography · CPC title

• G01N2035/00485

  combined with sample carriers · CPC title