CN216236988U

CN216236988U - High-throughput automatic protein transfection platform

Info

Publication number: CN216236988U
Application number: CN202122563151.6U
Authority: CN
Inventors: 徐婷婷; 刘鹏; 秦毅; 王卓智; 顾继杰; 陈智胜
Original assignee: Shanghai Yaoming Biomedical Co ltd
Current assignee: Shanghai Yaoming Biomedical Co ltd
Priority date: 2021-10-25
Filing date: 2021-10-25
Publication date: 2022-04-08
Anticipated expiration: 2031-10-25

Abstract

The utility model discloses a high-flux automatic protein transfection platform which comprises a workbench body and a special culture bottle base, wherein the special culture bottle base is fixedly arranged at the top of a table top of the workbench body, the special culture bottle base is used for fixing a 125mL culture bottle on the table top, an air filtering system is arranged at the top of the workbench body, a sliding rail is further arranged on the workbench body, a liquid treatment mechanical arm is slidably arranged on the sliding rail, and a liquid storage device, a liquid transfer gun head box, a twenty-four-hole deep hole plate, a twenty-four-hole freezing box, a vacuum extraction module and a test tube rack are further sequentially arranged on the table top of the workbench body at intervals. The utility model can automatically complete the transfection operation, simultaneously improves the transfection flux, saves the operation time and the labor cost compared with manual transfection, and simultaneously avoids the generation of experimental errors.

Description

High-throughput automatic protein transfection platform

Technical Field

The utility model relates to the technical field of biological medicines, in particular to a high-throughput automatic protein transfection platform.

Background

Transfection, which is a process in which a eukaryotic cell actively or passively introduces a foreign DNA fragment under certain conditions to obtain a new phenotype, is divided into transient transfection, in which the foreign DNA/RNA is not integrated into the host chromosome, and stable transfection, so that multiple copy numbers can exist in one host cell to generate high-level expression, but usually lasts for only several days, and is mostly used for analysis of promoters and other regulatory elements; stable transfection, also referred to as permanent transfection, where foreign DNA may be either integrated into the host chromosome or may exist as an episome, has become the fundamental method often involved in laboratory work as studies of gene and protein function have progressed.

In the biological science development stage, high-throughput transient expression of small-volume proteins is required to produce enough proteins to meet the requirements of protein screening. At present, most of all units and laboratories are manually operated by experimenters to perform protein expression transfection operation, and the whole process of protein transfection relates to a plurality of steps of plasmid filtration, transfection reagent configuration, transfection reagent and plasmid mixing, transfection reagent and plasmid mixture adding in cells and the like. For some laboratories, the specifications of protein transfection are fixed, but because the number of samples is large and the requirement on time period is high, namely the requirement on success rate is high, the requirements on operation proficiency and concentration of experimenters are extremely high, and when the experimental operation is difficult to reach the standard, the transfection is often affected, so that the flux is reduced or the accuracy is reduced.

Therefore, automation is an excellent solution for protein transfection with uniform specification, high flux and high accuracy requirement, automatic equipment of multiple companies such as Tecan, Beckman, HAMILTON, Thermo Fisher and the like is available on the market at present, Tecan free EVO 200 equipment is selected in the scheme, and the construction of a 40mL volume protein automatic transfection platform is completed.

SUMMERY OF THE UTILITY MODEL

In order to solve the technical problems, the utility model provides a high-throughput automatic protein transfection platform, which adopts the following technical scheme:

the utility model provides an automatic albumen transfection platform of high flux, includes the workstation body and the special base of blake bottle, the special base fixed mounting of blake bottle is in the top of the mesa of the workstation body, the special base of blake bottle is used for fixing 125mL blake bottle on the mesa, the top of workstation body is provided with filtration system, still be provided with the slide rail on the workstation body, slidable mounting has liquid treatment arm on the slide rail, still spaced in proper order on the mesa of workstation body be provided with reservoir, pipette gun head box, twenty-four holes deep hole board, twenty-four holes freezing box, vacuum extraction module and test-tube rack.

Specifically, the special base of blake bottle is provided with a plurality ofly, each the special base of blake bottle all includes base, bottom plate, silica gel board and fixed plate, base fixed mounting is at the top of the workstation body, bottom plate fixed mounting is at the top of base, the top of bottom plate is provided with a plurality of circular recesses, silica gel board fixed mounting is at the top of bottom plate, just seted up on the silica gel board a plurality ofly with the first through-hole of circular recess one-to-one, fixed plate fixed mounting is at the top of silica gel board, just set up on the fixed plate a plurality ofly with the second through-hole of first through-hole one-to-one.

Specifically, the air filtration system includes the laminar flow case, be provided with fan, prefilter and high efficiency filter in the laminar flow case, the laminar flow case is used for guaranteeing whole operational environment's aseptic condition.

Specifically, the special base of blake bottle is provided with three, circular recess is provided with eight.

Specifically, the high-throughput automated protein transfection platform further comprises a 40mL automated protein transfection expression program, wherein the 40mL automated protein transfection expression program is used for controlling the liquid treatment mechanical arm so that transfection can be automatically carried out.

At present, the protein transfection is mostly completed by adopting a manual operation mode in the existing protein expression process, the manual operation consumes time and labor for the high-flux transfection process, and the high-flux transfection process also has extremely high requirements on the operation accuracy and the concentration degree of operators in order to ensure high success rate.

Compared with the prior art, the utility model has the beneficial effects that:

1. due to the arrangement of the special base for the culture bottle, the Tecan workstation can be well adapted to the 125mL culture bottle, so that 40mL protein transfection can be carried out, the transfection flux is improved, and the transfection efficiency is improved;

2. through the setting of a 40mL automatic protein transfection expression program, a liquid treatment mechanical arm can be automatically controlled, and a plurality of operation steps such as plasmid filtration, transfection reagent preparation, transfection reagent and plasmid mixing, transfection reagent and plasmid mixture adding into cells and the like are completed;

3. the operation process steps of protein instantaneous transfection are many, and the sample is in large quantity, and under the condition that high flux transfection required, manual operation inevitably can produce the error, and when transfection flux was higher, the experimental error that the human factor arouses often can cause the waste of experimental time and material cost, and the degree of accuracy of automatic transfection is high, can effectively reduce the experimental error that the human factor arouses, reduces time and material cost that repeated experiment caused simultaneously.

Drawings

FIG. 1 is a schematic view of the overall structure of the present invention;

FIG. 2 is a schematic structural view of a base dedicated to a culture flask according to the present invention;

FIG. 3 is a flow chart of a 40mL automated protein transfection expression procedure of the present invention;

in fig. 1-2, 1, a table body; 2. a base special for a culture bottle; 21. a base; 22. a base plate; 23. a silica gel plate; 24. a fixing plate; 25. a circular groove; 3. an air filtration system; 4. a slide rail; 5. a liquid handling robot arm; 6. a reservoir; 7. a pipette tip box; 8. a twenty-four hole deep hole plate; 9. a twenty-four hole freezer; 10. a vacuum extraction module; 11. a test tube rack; in fig. 3, the steps in the rectangular box are automation content.

Detailed Description

The technical solutions of the present invention will be described clearly and completely with reference to the accompanying drawings, it should be noted that, in the following description, the terms "front", "rear", "left", "right", "upper" and "lower" refer to the directions in fig. 1, the terms "bottom" and "top", "inner" and "outer" refer to the directions toward or away from a specific component geometry, respectively, and the terms "first" and "second" are used for descriptive purposes only and are not to be interpreted as indicating or implying relative importance or implicitly indicating the number of indicated technical features, "plurality" means two or more, and the terms "mounted", "connected", "fixed" and the like are to be interpreted broadly, for example, "connected" may be a fixed connection, a detachable connection, or an integral connection; "coupled" may be direct or indirect through an intermediary. The specific meanings of the above terms in the disclosed embodiments of the utility model can be understood by those of ordinary skill in the art as appropriate. It is to be understood that the described embodiments are only a part of the embodiments of the present invention, and that the materials, instruments, reagents, etc. used in the following embodiments are commercially available, unless otherwise specified, and that the technical means used in the embodiments, if not specifically specified, are conventional means well known to those skilled in the art.

Example 1

As shown in FIG. 1, a high-throughput automatic protein transfection platform comprises a workbench body 1 and a culture bottle special base 2, wherein the culture bottle special base 2 is fixedly installed at the top of a table top of the workbench body 1, a 125mL culture bottle is generally used for high-throughput protein transfection of 40mL and is not used as a transfection container in a Tecan workstation, and a 125mL culture bottle base matched with the Tecan workstation is not arranged, so that the 125mL culture bottle is fixed on the table top by using the culture bottle special base 2, and further transfection operation is facilitated.

The technical effect of high-flux protein transfection is achieved by the aid of the technical scheme that a Tecan workstation is selected for use on the workbench body 1, an air filtering system 3 is arranged at the top of the workbench body 1, the air filtering system 3 works through a regulating fan, air speed is kept within an ideal range, air cleanness of a working area is guaranteed, a sliding rail 4 is further arranged on the workbench body 1, a liquid treatment mechanical arm 5 is slidably mounted on the sliding rail 4, the liquid treatment mechanical arm 5 can be selectively provided with a fixed sample adding needle and a disposable sample adding needle, different channel numbers can be selected, and technical effects of high-flux protein transfection are achieved Liquid-transfering gun rifle head box 7, twenty-four hole deep hole board 8, twenty-four hole freezing box 9, vacuum extraction module 10 and test-tube rack 11 remove, are convenient for accomplish various complicated liquid-transfering and application of sample operations, have improved work efficiency.

As shown in fig. 2, a plurality of special bases 2 for culture bottles are provided, each special base 2 for culture bottles comprises a base 21, a bottom plate 22, a silica gel plate 23 and a fixing plate 24, the base 21 is fixedly installed on the top of the worktable body 1, the bottom plate 22 is fixedly installed on the top of the base 21, a plurality of circular grooves 25 are provided on the top of the bottom plate 22, the silica gel plate 23 is fixedly installed on the top of the bottom plate 22, a plurality of first through holes corresponding to the circular grooves 25 are formed in the silica gel plate 23, the fixing plate 24 is fixedly installed on the top of the silica gel plate 23, a plurality of second through holes corresponding to the first through holes are formed in the fixing plate 24, when the culture bottles are placed, the bottoms of the culture bottles pass through the first through holes and the second through holes and enter the circular grooves 25, the silica gel plate 23 is provided to play a role of anti-skid, so as to ensure that the culture bottles can be stably and firmly fixed in the circular grooves 25, the setting of fixed plate 24 prevents that the blake bottle from rocking at liquid treatment arm 5's removal in-process to lead to influencing the stable of transfection operation and going on, blake bottle special base 2 is provided with threely, and circular recess 25 is provided with eight, has guaranteed that the sample quantity who moves a round is 24, thereby has improved albumen transfection flux.

Air filtration system 3 includes the laminar flow case, be provided with the fan in the laminar flow case, prefilter and high efficiency filter, the air intake and the outside air intercommunication of fan, the air outlet and the prefilter intercommunication of fan, prefilter and inside and high efficiency filter's inside intercommunication, high efficiency filter's air outlet and the inside intercommunication of workstation body 1, the fan inhales the prefilter with the air inside, the air after the prefilter gets into high efficiency filter again, high efficiency filter sends out the clean air after filtering to work area again, the laminar flow case is used for guaranteeing whole operational environment's aseptic condition.

The high-throughput automatic protein transfection platform further comprises a 40mL automatic protein transfection expression program, wherein the 40mL automatic protein transfection expression program is used for controlling the liquid treatment mechanical arm 5, so that transfection can be automatically carried out, experimental errors are not easy to generate, and time and material cost caused by repeated experiments are reduced.

As shown in FIG. 3, the transfection procedure using the present invention is specifically performed as follows:

1) using an Expi 293expression system of Thermo Fisher to perform protein transient expression, preparing 24 plasmid samples and placing the plasmid samples in a test tube for later use;

2) adding a cell sample into a 125mL culture bottle, and then placing the 125mL culture bottle in a special base 2 for the culture bottle;

3) starting a 40mL automatic protein transfection expression program, controlling a liquid treatment mechanical arm 5 by the 40mL automatic protein transfection expression program, completing multiple steps of plasmid filtration, transfection reagent preparation, transfection reagent and plasmid mixing, transfection reagent and plasmid mixture adding into cells and the like, and transferring the mixed sample into a 125mL culture bottle in the step 2);

4) and (3) putting the 125mL culture bottle in the step 2) into a shaking table to finish the transfection operation.

From the steps, the high-throughput automatic protein transfection platform provided by the utility model can effectively improve transfection flux and further improve transfection efficiency, and through the setting of a 40mL automatic protein transfection expression program, the liquid treatment mechanical arm 5 can be automatically controlled, the special base 2 for the culture bottle can fix a 125mL culture bottle on the top of a Tecan workbench, and the liquid treatment mechanical arm 5 is matched to complete a plurality of operation steps of plasmid filtration, transfection reagent preparation, transfection reagent and plasmid mixing, transfection reagent and plasmid mixture adding into cells and the like, the automatic program runs for one round, the number of samples capable of completing transfection operation is 24, the operation time is 60min, manual operation is not needed, the operation time of transfection is reduced, the labor cost is saved, the accuracy of automatic transfection is high, and experimental errors caused by human factors can be effectively reduced, and simultaneously, the time and material cost caused by repeated experiments is reduced.

In summary, the above-mentioned embodiments are merely preferred embodiments of the present invention, and are not intended to limit the scope of the present invention, and all other embodiments obtained by those skilled in the art without any inventive work based on the embodiments of the present invention shall fall within the scope of the present invention, and any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the scope of the present invention.

Claims

1. The utility model provides a high throughput automation protein transfection platform, includes the special base of workstation body (1) and blake bottle (2), its characterized in that: the utility model discloses a laboratory glassware, including workstation body (1), the top of the mesa of workstation body (1), the special base of culture flask (2) are used for fixing 125mL culture flask on the mesa, the top of workstation body (1) is provided with air filtration system (3), still be provided with slide rail (4) on the workstation body (1), slidable mounting has liquid treatment arm (5) on slide rail (4), still spaced in proper order on the mesa of workstation body (1) be provided with reservoir (6), liquid transfer rifle head box (7), twenty-four hole deep hole board (8), twenty-four hole freezing box (9), vacuum extraction module (10) and test-tube rack (11).

2. A high throughput automated protein transfection platform according to claim 1 wherein: the utility model discloses a culture flask, including base (21), bottom plate (22), silica gel plate (23), fixed plate (24), base (21) fixed mounting is at the top of workstation body (1), bottom plate (22) fixed mounting is at the top of base (21), the top of bottom plate (22) is provided with a plurality of circular recess (25), silica gel plate (23) fixed mounting is at the top of bottom plate (22), just seted up on silica gel plate (23) a plurality of with the first through-hole of circular recess (25) one-to-one, fixed plate (24) fixed mounting is at the top of silica gel plate (23), just set up on fixed plate (24) a plurality of with the second through-hole of first through-hole one-to-one.

3. A high throughput automated protein transfection platform according to claim 1 wherein: the air filtering system (3) comprises a laminar flow box, wherein a fan, a pre-filter and a high-efficiency filter are arranged in the laminar flow box, and the laminar flow box is used for ensuring the sterile condition of the whole operation environment.

4. A high throughput automated protein transfection platform according to claim 2 wherein: the special base (2) for the culture bottle is provided with three, and the circular grooves (25) are provided with eight.

5. A high throughput automated protein transfection platform according to claim 1 wherein: the high-throughput automated protein transfection platform further comprises a 40mL automated protein transfection expression program, wherein the 40mL automated protein transfection expression program is used for controlling the liquid treatment mechanical arm (5) so that transfection can be automatically carried out.