CN205374079U - Bioprocess rapid sampling device - Google Patents
Bioprocess rapid sampling device Download PDFInfo
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- CN205374079U CN205374079U CN201520912165.6U CN201520912165U CN205374079U CN 205374079 U CN205374079 U CN 205374079U CN 201520912165 U CN201520912165 U CN 201520912165U CN 205374079 U CN205374079 U CN 205374079U
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Abstract
The utility model relates to a bioprocess rapid sampling device, it includes sampling tube, valve, peristaltic pump, still include time schedule controller better. The utility model discloses a fermentation process sample collection and cancellation in 1 second can be realized to the device, have improved bioprocess data acquisition's accuracy and reliability greatly. Has simple structure simultaneously, characteristics that the suitability is strong.
Description
Technical field
This utility model belongs to biological sampling devices field, more specifically, this utility model relates to a kind of bioprocess rapid sampling attachment.
Background technology
For bioprocess investigated carry out multiparameter on-line checking (environment pH, dissolved oxygen, speed of agitator, exhaust gas composition etc.) outward, also need to the outer intracellular organic matter concentration of the various born of the same parents of determined off-line, bioprocess to be carried out the biological study of system, verify the mechanism of bioprocess.But, the detection for intracellular organic matter often also exists various problems so that it is the reliability of detection parameter enjoys query with accuracy.
One of them subject matter is to change because of the change of environment at sample thalline self after leaving yeasting, thus losing its representativeness during the fermentation.Such as, after leaving the bioreactor of continuous stirring and ventilation, the thalline of high concentration consumes rapidly dissolved oxygen in the sample due to oxygen process, hence into strict anaerobic processes.If now still having remained part glucose not utilized (residual sugar) in fermentation liquid, then thalline can consume this part sugar source by anaerobic pathway and produce alcohols and organic acid etc..This makes the sugared concentration of late detection be underestimated on the one hand, the balance of intracellular organic matter also can be made to change simultaneously.All above-mentioned changes are all likely within the time quickly to occur: the sugared concentration reduction outside born of the same parents can detect in several minutes, and the reduction that the ATP energy level in born of the same parents causes because of oxygen restriction was likely to occur in the several seconds.
For problems, designed both at home and abroad and apply multiple rapid sampling attachment.It is illustrated in figure 1 two kinds of rapid sampling attachments of same type, it is achieved that the continuous fast sampling within the short time (several minutes).It is hardware foundation that this sampling mode generally requires by comparatively large-scale complete instrument and the accurate sampling pump controlled.
It is illustrated in figure 2 another kind of rapid sampling attachment continuously, and adds disturbed conditions wherein, catch to realizing thalline dynamic response under disturbed conditions.
Fig. 3 is the third rapid sampling attachment, and this device produces negative pressure by vacuum pump and the speed of sampling is accelerated, it is achieved the second level fast sampling of stricti jurise.
A few class rapid sampling attachments achieve the fast sampling process of bioprocess all to a certain extent above.But they are all respectively arranged with shortcoming.
Rapid sampling attachment as shown in Figure 1, its structure is complicated, the specificity of system is strong, cannot simply overlap for all kinds of reactors, simultaneously because be directed to multiple Automated condtrol, harmony between system and system needs instruction, and the time point of continuous sampling is more difficult accurate control also, and one is sized also more difficult amendment.
Rapid sampling attachment as shown in Figure 2, trades space for time, it is possible to accurately estimates sampling time point by calculating flow velocity with distance, fixes yet with its sample position, it is impossible to arbitrarily according to experiment Demand Design sampling process.And, front two class reactors are subject to the restriction of sampling caliber (sampling dead volume) and the fast sampling that often cannot realize in 1 second and cancellation process, and cannot be properly functioning when running into high viscosity fermentation (filamentous fungi fermentation and polysaccharide fermentation).The weighing process offed normal both had added the time of sampling process, also can introduce the error of sampling amount because of low temperature quencher condensation.
Rapid sampling attachment as shown in Figure 3, builds negative pressure space by adding extra vacuum pump, it is achieved that the fast sampling process within 1 second.But, whole system still also exists the shortcomings such as the suitability not strong (needing to specify the supporting test tube adapter of test tube), system reliability not strong (negative pressure space tends to lose efficacy, and causes sampling unsuccessfully) because of gas leakage.
Therefore, this area also needs to the rapid sampling attachment of the improvement that development structure simplifies, sampling is accurate, the suitability is strong.
Utility model content
The purpose of this utility model is in that to provide a kind of bioprocess rapid sampling attachment.
In first aspect of the present utility model, it is provided that a kind of biological respinse liquid (fermentation liquid) sampler, described device includes:
Probe tube (1), it is T tube, including the first sampling section (11), the second sampling section (12), waste liquid section (13)；
First valve (2), it is arranged in the first sampling section (11), can control connection and the Guan Bi of the first sampling section (11)；
Second valve (3), it is arranged in the second sampling section (12), can control connection and the Guan Bi of the second sampling section (12)；
Peristaltic pump (4), it is arranged in waste liquid section (13), can control sampling before drain waste liquid flow velocity and control sampling after device emptying.
In a preference, described bioprocess rapid sampling attachment includes:
Time schedule controller (5), it is connected with described the first valve (2), the second valve (3) and peristaltic pump (4), controls the first valve (2), being turned on and off of the second valve (3) and peristaltic pump (4).
In another preference, in described bioprocess rapid sampling attachment, the first described valve and the second valve be: pipe clip valve.
In another preference, described bioprocess rapid sampling attachment also includes:
First container (6), is used for receiving the waste liquid that waste liquid section (13) flows out.
In another preference, described bioprocess rapid sampling attachment also includes:
Second container (7), is used for receiving the sample that the second sampling section (12) flows out.
In another preference, in described bioprocess rapid sampling attachment, described second container (7) is equipped with fermentation liquid cancellation solution.
Other side of the present utility model, due to this disclosure, is apparent to those skilled in the art.
Accompanying drawing explanation
Fig. 1, the continuous rapid sampling attachment of multiple spot of the prior art.
Fig. 2, the continuous rapid sampling attachment with disturbance experiments point of the prior art.
Fig. 3, single-point rapid sampling attachment of the prior art.
Single-point rapid sampling attachment structural representation in Fig. 4, embodiment 1.
Fig. 5, rapid sampling attachment collimation are tested.A, C, E:250ms sample time；B, D, F:500ms sample time；A, B: probe tube internal diameter 3.1mm；C, D: probe tube internal diameter 2.4mm；E, F: probe tube internal diameter 3.1mm.
Average sample number under Fig. 6, different reactor pressure (0.03MPa, 0.04MPa, 0.05MPa), sample time (250ms, 500ms) and probe tube specification (internal diameter 1.6mm, 2.4mm, 3.1mm).
Wherein, in accompanying drawing, each numeral description of symbols is as follows:
1: probe tube；
11: the first sampling sections, 12: the second sampling sections；
13: waste liquid section；
2: the first valves, 3: the second valves；
4: peristaltic pump；
5: time schedule controller；
6: the first containers, 7: second container；
100: pipe box；
101: pipe clip valve；
102: gear band；
103: stepping engine；
104: rotary valve dish；
105: stepper motor；
106: three-way valve；
107: three-way valve；
108: pipe clip valve I；
109: pipe clip valve II；
110: tube regulator；
111: waste liquid；
112: Dewar vessel；
113: developmental tube.
Detailed description of the invention
In the process utilizing bioreactor, in order to realize sampling fast and efficiently, the design people have passed through and in depth studies, it is provided that a kind of sweat rapid sampling attachment based on bioreactor.Device of the present utility model is capable of the sweat sample collecting in 1 second and cancellation, substantially increases accuracy and the reliability of bioprocess data acquisition.There is simple in construction simultaneously, the feature that the suitability is strong.
The probe tube of device of the present utility model is a kind of T tube, and this T tube includes several sampling section, each sampling section arranges valve or peristaltic pump, controls the flow direction of sample liquid.
Described T tube can be prepared by multiple material, and this utility model has no particular limits for its material used, for instance elastomeric material, silica gel material.As optimal way of the present utility model, described T tube is silica gel tube.
The caliber of described T tube can be determined according to actual sampling scale, has no particular limits in this utility model.As optimal way of the present utility model, the interior caliber of described T tube can be 1-5mm；Preferably 1.5-3mm, for instance 1.5,2,2.5,3mm.Certainly, the scale according to actual bioreactor scale and sampling, its caliber can also exceed this scope, as long as guaranteeing to meet the fast sampling standard completing a complete set of flow process in 1s.
As optimal way of the present utility model, described valve is pipe clip valve；It is preferred that it is normally closed pipe clip valve.Pipe clip valve controls connection and the Guan Bi of the pipeline section at its place.
Peristaltic pump described in the utility model is arranged in the waste liquid section of T-shaped pipe, and it can control probe tube waste discharge and emptying.As optimal way of the present utility model, described peristaltic pump is high speed peristaltic pump, it is possible to form the rotating speed be more than or equal to 5ml/s.
Device of the present utility model can complete discharging of waste liquid in the pipe in special time (namely avoiding the interior remaining waste impact on sample of previously pipe) by the running of peristaltic pump, when peristaltic pump decommissions, and can by bioreactor internal positive pressure power (> 0.03MPa) complete fast sampling process.
As optimal way of the present utility model, time schedule controller is adopted to be connected with described valve and peristaltic pump, being turned on and off of valve described in control and peristaltic pump.Described time schedule controller can adopt time schedule controller known in the art, drives different valves and peristaltic pump to carry out starting or closing according to rule by the formulation time.Time schedule controller has simple human-computer interaction interface, it is possible to change opening and closing operations and the persistent period of each valve and peristaltic pump according to experiment demand in good time.
Bioprocess rapid sampling attachment of the present utility model realizes quantitative quick sampling effectively by simple hardware device (silica gel tube, pipe clip valve, peristaltic pump, time schedule controller), and has good system compatibility.
Bioprocess rapid sampling attachment of the present utility model designs based on feature succinct, reliable, that the suitability is strong, realize fast sampling in 1 second under minimal hardware, and introduce the sampling line overlapping different tube diameters more, improve the compatibility for different fermentations process (low high viscosity).
Bioprocess rapid sampling attachment of the present utility model also can realize thalline cancellation simultaneously, and the accurate quantitative analysis for materials various in thalline born of the same parents provides hardware foundation.
Compared to current existing rapid sampling attachment, the beneficial effect of bioprocess rapid sampling attachment of the present utility model includes:
1. simple in construction, it is easy to safeguard, can adaptive multiple sampling line, for the fast sampling of the kinds of experiments room scale bioreactors such as 250ml, 1.5L, 5L, 7L, 15L, 30L, 50L；
2. specific sample program can be set, it is achieved the automated procedure rinsing, sampling, rinse.Different flush volume and sample volume can be realized by regulating time schedule controller parameter；Can directly regulate sample time to control accurate sampling amount；
3. sampling amount has good collimation；
4. can reach the fast sampling standard of gram level in 1 second；
5. can complete samples weighing in sampling simultaneously, reduce error of sampling.
Below in conjunction with specific embodiment, this utility model is expanded on further.Should be understood that these embodiments are merely to illustrate this utility model rather than limit scope of the present utility model.The experimental technique of unreceipted actual conditions in the following example, generally conventionally condition such as J. Pehanorm Brooker etc. are write, Molecular Cloning: A Laboratory guide, the third edition, Science Press, the condition described in 2002, or according to manufacturer it is proposed that condition.
Embodiment 1, bioprocess rapid sampling attachment
1, device 1
Described bioprocess rapid sampling attachment includes:
Probe tube 1, it is T tube, including the first sampling section 11, the second sampling section 12, waste liquid section 13；Three mouth of pipe ends of T tube are respectively provided with opening port.This probe tube 1 is silica gel tube, and its internal diameter is 2.4mm.
Pipe clip valve 2 (normally closed), it is arranged in the first sampling section 11, can control connection and the Guan Bi of the first sampling section 11；
Pipe clip valve 3 (normally closed), it is arranged in the second sampling section 12, can control connection and the Guan Bi of the second sampling section 12；
Peristaltic pump 4 (normally closed), it is arranged in waste liquid section 13, can control sampling before drain waste liquid flow velocity and sampling after device emptying.This peristaltic pump 4 is high speed peristaltic pump, it is possible to form the rotating speed be more than or equal to 5ml/s.
Time schedule controller 5 (single-chip microcomputer), it is connected with described pipe clip valve 2, pipe clip valve 3 and peristaltic pump 4, controls opening or closing of pipe clip valve 2, pipe clip valve 3 and peristaltic pump 4.Opening and closing by the clock control chip controls relay in time schedule controller 5.Pipe clip valve is opened, then the corresponding pipeline section connection of probe tube, and pipe clip valve is closed, then the corresponding pipeline section Guan Bi of probe tube.
First container 6, for receiving the waste liquid that waste liquid section 13 flows out；
Second container 7, for receiving the sample that the second sampling section 12 flows out.
2, device 2
Assembling a bioprocess rapid sampling attachment, its silica gel tube internal diameter is 3.1mm, other parts and arrange same device 1.
3, device 3
Assembling a bioprocess rapid sampling attachment, its silica gel tube internal diameter is 1.6mm, other parts and arrange same device 1.
Embodiment 2, bioprocess rapid sampling attachment Working Examples
The present embodiment relates to being sampled for the cell fermentation liquid in bioreactor (fermentation tank).Sampling process is as follows:
1. prepare suitable quencher in advance and be stored on request in temperature chamber.
2. sampling parameters is set by time schedule controller: (1) scavenging period 1, gives tacit consent to 1000 milliseconds；(2) scavenging period 2, give tacit consent to 700 milliseconds；(3) sample time, 250 milliseconds are given tacit consent to；(4) peristaltic pump rotating speed, gives tacit consent to maximum (top) speed 600RPM.
3. before sampling, the probe tube containing quencher is placed in electronic balance, zero setting by the several seconds.
4. the opening that first samples section 11 is inserted in bioreactor.Start sampling, run sampling by pre-set programs:
(1) pipe clip valve 11 and peristaltic pump 4 are opened, peristaltic pump acquiescence rotating speed 600RPM.Now fermentation liquid is sampled section 11 by first and flows into waste liquid bottle (the first container 6) through waste liquid section 13.This stage is used for cleaning the first sampling section 11, the remaining fermentation liquid in the pipeline of waste liquid section 13, with emptying before fermentation liquid, guarantee sample time obtain fermentation liquid for sample in actual tank.
(2) step (1) continues 1000 milliseconds (scavenging period 1).
(3) closing peristaltic pump, open pipe clip valve 11 and pipe clip valve 12, be sampled by bioreactor internal positive pressure power, sample liquid flows into second container 7, completes sampling process.
(4) step 3 continues 250 milliseconds (sample time).
(5) close pipe clip valve 11, open pipe clip valve 12 and peristaltic pump 4, evacuate and remain in the second sampling section 12, the fermentation liquid that waste liquid section is 13 sections interior.
(6) step 5 continues 700 milliseconds (scavenging period 2).
(7) sampling terminates.
5. rapid recorded electronic balance reading after having sampled, and continue the steps such as cell filtration subsequently, intracellular organic matter extracting.
Embodiment 3, bioprocess rapid sampling attachment Performance Evaluation
1, sampling collimation
Choose 3 kinds of sampling silica gel tubes (internal diameter is 3.1mm, 2.4mm and 1.6mm respectively) that embodiment 1 makes respectively.For sample time, it is respectively provided with 2 kinds of sample times: 250 milliseconds and 500 milliseconds.With pure water for fluid, test the sampling amount parallel lines of continuous 10 sub-samplings of this system.
Result is as shown in Figure 5.All experimental results all show good system collimation, and error of sampling is all within 5%.
2. sampling amount and retention time
Experiments verify that, the probe tube specification that the actual sampling amount of this rapid sampling attachment operates overvoltage, sample time and use with reactor is relevant, unrelated with ventilation, speed of agitator etc..Specific experiment result is as shown in Figure 6.
Table 1, different probe tube specification and the flow velocity under reactor pressure and retention time
Except carrying out specifying the sampling work of quality exactly, rapid sampling attachment also needs to ensure being rapidly completed of whole sampling process.Standards of measurement are in fermentation liquid retention time in whole sampler.Its calculation is:
Retention time [s]=(in reactor dead volume [ml]+sampler dead volume [ml])/(total sampling amount [ml]/sample time [s])
The retention time of this rapid sampling attachment is as shown in table 1.It will be seen that under multiple conditional combination, its retention time is respectively less than 1 second, has reached the standard of fast sampling.
3. the application of real attenuation process and data result
This rapid sampling attachment has been applied in the sweat sampling of aspergillus niger (Aspergillusniger) and Penicllium chrysogenum (Penicilliumchrysogenum).Shown in sampling actual effect such as table 2 and table 3:
The continuous fast sampling sampling amount log of table 2, fermentation of Aspergillus niger process
Different fermentations stage fast sampling log in table 3, Penicllium chrysogenum sweat
Wherein, table 2 demonstrates in real attenuation process, for single fermentation time point, this rapid sampling attachment can time continuous fast sampling quickly and accurately, the error between each sample point is less than 5%.
Table 3 illustrates the different phase at sweat, and sampling amount can be gradually lowered along with the increase of fermentation liquid bacterium amount, and this increase with filamentous fungi apparent viscosity during the fermentation is proportionate.In real attenuation process can by regulate sample time and carry out before sample point examination sampling guarantee that sampling amount is required sampling amount.
Should be understood that this utility model can be made various changes or modifications by those skilled in the art after having read above-mentioned teachings of the present utility model, these equivalent form of values fall within the application appended claims limited range equally.
Claims (6)
1. a bioprocess rapid sampling attachment, it is characterised in that described device includes:
Probe tube (1), it is T tube, including the first sampling section (11), the second sampling section (12), waste liquid section (13)；
First valve (2), it is arranged in the first sampling section (11), can control connection and the Guan Bi of the first sampling section (11)；
Second valve (3), it is arranged in the second sampling section (12), can control connection and the Guan Bi of the second sampling section (12)；
Peristaltic pump (4), it is arranged in waste liquid section (13), can control sampling before drain waste liquid flow velocity and control sampling after device emptying.
2. bioprocess rapid sampling attachment as claimed in claim 1, it is characterised in that described device includes:
Time schedule controller (5), it is connected with described the first valve (2), the second valve (3) and peristaltic pump (4), controls the first valve (2), being turned on and off of the second valve (3) and peristaltic pump (4).
3. bioprocess rapid sampling attachment as claimed in claim 1, it is characterised in that described the first valve (2) and the second valve (3) be: pipe clip valve.
4. bioprocess rapid sampling attachment as claimed in claim 1, it is characterised in that described device also includes:
First container (6), is used for receiving the waste liquid that waste liquid section (13) flows out.
5. bioprocess rapid sampling attachment as claimed in claim 1, it is characterised in that described device also includes:
Second container (7), is used for receiving the sample that the second sampling section (12) flows out.
6. bioprocess rapid sampling attachment as claimed in claim 5, it is characterised in that described second container (7) is equipped with fermentation liquid cancellation solution.
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| CN201520912165.6U CN205374079U (en) | 2015-11-16 | 2015-11-16 | Bioprocess rapid sampling device |
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| CN201520912165.6U CN205374079U (en) | 2015-11-16 | 2015-11-16 | Bioprocess rapid sampling device |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106840767A (en) * | 2017-01-17 | 2017-06-13 | 杭州泰林精密仪器有限公司 | A kind of sampling filtering device |
| CN107621387A (en) * | 2017-09-12 | 2018-01-23 | 河南科技大学第附属医院 | A kind of medical test sampler |
-
2015
- 2015-11-16 CN CN201520912165.6U patent/CN205374079U/en active Active
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106840767A (en) * | 2017-01-17 | 2017-06-13 | 杭州泰林精密仪器有限公司 | A kind of sampling filtering device |
| CN107621387A (en) * | 2017-09-12 | 2018-01-23 | 河南科技大学第附属医院 | A kind of medical test sampler |
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