CN108957020A - AKTA automatic sampling tomographic system and its application - Google Patents

Abstract

The invention discloses a kind of AKTA automatic sampling tomographic system, which includes the AKTA tomography devices interconnected and autosampler.The present invention is by connecting high performance autosampler with AKTA tomography devices common in protein purification, realize high throughput, the full-automation purifying of albumen, not only greatly improve working efficiency, instrument utilization rate and screening/production capacity to platform, and reduce manpower intervention, human resources are saved, manual operation bring is reduced and potentially makes mistakes risk.

Description

AKTA automatic sampling tomographic system and its application

Technical field

The present invention relates to biological fields, purify more particularly to the high throughput of protide large biological molecule, more specifically, Be be related to a kind of AKTA automatic sampling tomographic system, and its small size albumen high throughput purifying in application.

Background technique

Now, monoclonal antibody or other kinds of protide biopharmaceutical macromolecular drug have become a kind of very important life Object molecule, the research for clinical treatment or pathogenesis.But the research of exploitation or molecular function for a kind of protein drug, The purifying and preparation that hundreds of candidate molecules carried out with milligram grade or more are generally required, suitable target molecule is filtered out, therefore, Purifying flux is a key factor for realizing quickly screening suitable targets molecule, is quick preparation various types albumen and its number Measure the bottleneck of numerous derivatives.

The advantages of equipment currently used for purifying protein is the AKTA tomographic system of GE, this tomographic system can be achieved on The complicated purifications such as more chromatographic enrichment, continuous gradient elution, and chromatogram real time monitoring may be implemented.The disadvantage is that this tomographic system Sampling valve is limited, removes buffer channel, once can only at most run seven samples, greatly limits flux and the work of purifying Efficiency.For purifying multiple 20mL samples, conventional method can only 7 samples of each run, then manual flushing sample channel, Pipeline is correspondingly placed into next group sample again, intricate operation and be easy to cause mistake, the time is long also to generate product quality It influences.

Existing high-throughput purifying is usually to be completed using the pipette tips or porous plate of liquid working station collocation prepackage filler, The advantages of such way of purification is: 1. flux are high, can disposably complete the purifying of 96 samples；2. the operation in microwell plate It is easier to be connected with the high throughput analysis in downstream.But disadvantage is similarly obvious: the purifying of 1. singles is small in size, usually in 2mL Left and right, purified product tend not to the analysis detection requirement for meeting the later period；2. purification mode is simple, be typically only capable to be a step parent And purifying；3. lacking the real time monitoring of chromatogram.

There is a autosampler at present, its sample pipe support can disposably put 240 according to the difference of volume, maximum A sample is very suitable for the automation purifying of multiple samples, current purifying flux can be greatly improved.But such instrument is usual It is for being combined some spectrometric instruments, instrument producer records without the combination with AKTA tomography devices, between two instrument Communication cooperation problem before this without solution.

Summary of the invention

One of the technical problem to be solved in the present invention is to provide a kind of AKTA automatic sampling tomographic system, its stability and Adaptability is high, can greatly improve protein purification automation flux.

In order to solve the above technical problems, AKTA automatic sampling tomographic system of the invention, the AKTA layer including interconnection Desorption device and autosampler.

The tomography devices are GE-AKTA series tomography devices, and the autosampler is that Teledyne CETAC is automatic Sample injector.

The autosampler is connect by connecting line with the AKTA tomography devices.

Optionally, when the tomography devices are GE-AKTA Pure, the AKTA Pure passes through I/O Box E9 and institute Autosampler connection is stated, and realizes the auto-flushing of sample introduction pipeline after loading by designing additional flow path.

Optionally, the order of one group of transmission signal is encoded in the operating software of the AKTA tomography devices, it is corresponding, It is encoded with the order for waiting this signal in the operating software of the autosampler, the collaborative work of two instruments is realized with this.

The sample volume of the AKTA automatic sampling tomographic system is preferably 2ml-150mL.

The present invention is by by common AKTA in high performance Teledyne CETAC autosampler and protein purification Tomographic system connection, the high throughput for realizing 2-150mL volume of protein sample, full-automatic purifying are (compared to AKTA, flux peak Improve 30 times or more), working efficiency, instrument utilization rate and screening/production capacity to platform are not only greatly improved, but also Reduce manpower intervention, save human resources, reduces manual operation bring and potentially make mistakes risk.

Detailed description of the invention

Fig. 1 is the schematic wiring diagram of autosampler I/O connector.

Fig. 2 is the schematic wiring diagram of the I/O Box E9Digital in/out interface of AKTA system.

Fig. 3 is the structural schematic diagram of AKTA automatic sampling tomographic system of the invention.

Fig. 4 is that 3 duplicate chromatograms of each sample compare in embodiment.Wherein, A~F figure respectively corresponds number and is 1222 (1-3), 1223 (1-3), 1225 (1-3), 1229 (1-3), 1231 (1-3), 1219 (1-3) antibody samples.

Specific embodiment

To have more specific understanding to technology contents of the invention, feature and effect, now in conjunction with the drawings and specific embodiments, Technical solution of the present invention is further described in detail.

The configuration and purifying of embodiment 1AKTA automatic sampling tomographic system

The configuration of the AKTA automatic sampling tomographic system of the present embodiment and purification process are as follows:

Step 1: configuration AKTA system is used to connect the I/O Box E9 (GE-29011361) of external equipment, and is used for Connect the connecting line of autosampler and I/O Box E9.

Step 2: installation autosampler, and the computer that AKTA Pure equipment connects is connected to by included interface.

Step 3: installation I/O Box E9, V9 (H)-V to AKTA Pure equipment.

Step 4: the sample introduction needle of autosampler being connected to AKTA IS valve, AKTA IS valve is connected to V9 (H) valve, V9 (H) valve connects sample pump intake, and AKTA Outlet valve connects V9 (H) valve.All other routes are constant by default connection.This connection mesh : switched using autosampler control sample introduction needle in different sample rooms, pumps control loading using AKTA sample, utilize V9 (H) sample introduction needle inner wall is cleaned by back flush after one Sample Purification on Single of valve realization.

Step 5: being connected autosampler with AKTA Pure I/O Box E9 by connecting line.

Step 6: being write in the operating software AScript of the operating software UNICORN of AKTA and autosampler respectively Method program.

The difference of AKTA method and original method is: 1. after completion of the sample AKTA have the order for issuing signal, it is corresponding , there is the order for waiting this signal in the specific node of AScript method, the collaborative work of two instruments is realized with this.2. After purifying terminates, the order of sample introduction needle is rinsed in addition.3. after having rinsed sample introduction needle, closing pump in time, is issued and believed by AKTA So that autosampler is received sample introduction needle and rinses the instruction completed.

AScript method is write referring to the explanation and purification requirements in random CD.

Step 7: preparing sample to be purified according to a conventional method, sample opening is put in order and is placed in sample pipe support.First It runs the AScript method write and runs the side UNICORN write after being moved in first sample to sample introduction needle Method.Check waits program automatic running to terminate after there is no problem.

The verifying of 2 reliability and stability of embodiment

Below by way of the antibody samples of 18 20mL volumes of purifying, the AKTA automatic sampling of the embodiment of the present invention 1 is chromatographed The reliability and stability of system are verified.Specific purification step are as follows:

(1) antibody samples number is 1222_1,1222_2,1222_3,1223_1,1223_2,1223_3,1225_1, 1225_2、1225_3、1229_1、1229_2、1229_3、1231_1、1231_2、1231_3、1219_1、1219_2、1219_3。 These samples are expressed in Chinese hamster ovary celI, are taken its fermentation liquid, are taken supernatant after centrifugal filtration.This 18 20ml supernatants are filled respectively Enter 50ml centrifuge tube, then this 18 50ml centrifuge tubes are disposably placed in order on the pipe support of autosampler.

(2) it conventionally carries out disinfection to AKTA system, to be used later with pure water and corresponding buffer solution for cleaning All pipelines.

(3) sample introduction needle outer wall is cleaned with the cleaning procedure that autosampler carries.

(4) pass through AKTA Program Purge sample introduction needle inner wall.

(5) the AScript method write is run, is completed after cleaning and being inserted into first sample to sample introduction needle, operation AKTA method.

(6) program operation is waited to complete.It collects sample and surveys final purification yield, as a result as shown in table 1 and Fig. 4, the result It demonstrates this AKTA automatic sampling tomographic system and is disposably purifying the reliability and stability in multiple samples.

The purification yield of 1 18 20ml antibody samples of table

Claims (6)

1.AKTA automatic sampling tomographic system, which is characterized in that AKTA tomography devices and autosampler including interconnection.

2. AKTA automatic sampling tomographic system according to claim 1, which is characterized in that the tomography devices are GE- AKTA tomography devices；The autosampler is Teledyne CETAC autosampler.

3. AKTA automatic sampling tomographic system according to claim 2, which is characterized in that the autosampler passes through company Wiring is connect with the AKTA tomography devices.

4. AKTA automatic sampling tomographic system according to claim 3, which is characterized in that the behaviour of the AKTA tomography devices Make software by carrying out signal communication with the operating software of the autosampler, realizes the collaborative work of two kinds of instruments.

5. AKTA automatic sampling tomographic system according to claim 1, which is characterized in that the AKTA automatic sampling chromatography The sample volume of system is 2ml-150mL.

6. any one of claim 1-5 AKTA automatic sampling tomographic system the answering in protide biological macromolecule purifying With.