CN107056879A - Multichannel Solid-phase synthesis peptides device - Google Patents
Multichannel Solid-phase synthesis peptides device Download PDFInfo
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- CN107056879A CN107056879A CN201710135871.8A CN201710135871A CN107056879A CN 107056879 A CN107056879 A CN 107056879A CN 201710135871 A CN201710135871 A CN 201710135871A CN 107056879 A CN107056879 A CN 107056879A
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- nitrogen
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- cylinder body
- adapter
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/04—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length on carriers
- C07K1/045—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length on carriers using devices to improve synthesis, e.g. reactors, special vessels
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/04—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length on carriers
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/04—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length on carriers
- C07K1/047—Simultaneous synthesis of different peptide species; Peptide libraries
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/06—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length using protecting groups or activating agents
- C07K1/061—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length using protecting groups or activating agents using protecting groups
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/06—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length using protecting groups or activating agents
- C07K1/08—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length using protecting groups or activating agents using activating agents
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- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Biochemistry (AREA)
- Biophysics (AREA)
- Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Analytical Chemistry (AREA)
- Peptides Or Proteins (AREA)
Abstract
The invention discloses a kind of multichannel Solid-phase synthesis peptides device, including reaction cylinder body, reaction cylinder base is connected with suction filtration adapter and nitrogen adapter, and nitrogen adapter is connected with nitrogen flexible pipe, and nitrogen adapter is provided with nitrogen valve, and suction filtration adapter is provided with suction filtration valve；Suction filtration adapter, which is connected with, vacuumizes flexible pipe；React and gas inlet/outlet pipe is provided with the top of cylinder body, reaction cylinder deck is connected with some connection grounds, each connection ground and is inserted with a Peptide systhesis pipe respectively, the bottom of the bottom intercalation reaction cylinder body of Peptide systhesis pipe and provided with core filter core；The middle part of Peptide systhesis pipe is provided with frosted connecting portion.The present invention carries out batch reaction by the way that different Peptide systhesis pipes is inserted into same reaction cylinder body simultaneously, and the efficiency compared with the past for substantially increasing Solid-phase synthesis peptides has saved the time of corresponding research, being capable of a variety of derived peptides of Fast back-projection algorithm.
Description
Technical field
The present invention relates to a kind of chemical experiment device, more particularly to a kind of Solid-phase synthesis peptides device.
Background technology
Solid-phase synthesis peptides are that protected amino acid is connected on solid resin, slough blocking group, then will be next
The residue of protected amino acid and first amino acid reacts and forms peptide bond, then sloughs the blocking group of second amino acid,
Next amino acid is reconnected, so circulation goes down to increase peptide chain.In Solid-phase synthesis peptides, solid resin must be used
Reaction substrate.
, it is necessary to using Suction filtration device in Solid-phase synthesis peptides reaction, Suction filtration device include bottle,suction and with bottle,suction phase
The vavuum pump of connection.
In Solid-phase synthesis peptides, the reaction solution for dissolving in activator is Coupling Solution, and the reaction solution for dissolving in deprotection agent is de-
Protect solution.When carrying out peptide solid phase synthesis, usually using Solid-phase synthesis peptides pipe, added in Solid-phase synthesis peptides pipe
Resin, first by resin swelling, is then alternately added deprotection solution and Coupling Solution carries out synthetic reaction.Add deprotection solution
When, resin with deprotection solution slough the reaction of blocking group；When adding Coupling Solution, resin and the band in Coupling Solution
The amino acid of blocking group carries out coupling reaction.
In Solid-phase synthesis peptides, it is often necessary to based on a female peptide, the different derivative in synthesis various mutations site
Peptide, such as when researching and developing polypeptide drug, it is necessary to based on a kind of female peptide, the different derived peptide in synthesis various mutations site, so
The optimal polypeptide drug of drug effect is found from various derived peptides afterwards.
An amino acid is often connected on resin, is required for being adapted using a kind of amino acid sequence specifically with polypeptide
Coupling Solution, connect different amino acid and need to use different Coupling Solutions.Therefore, in the process of Solid-phase synthesis peptides
In, it is necessary to repeatedly change reaction solution.When preparing a variety of derived peptides based on a kind of female peptide, the major part that various derived peptides need
Coupling Solution is identical, but current Solid-phase synthesis peptides device, identical in the Coupling Solution for preparing each derived peptide needs
When, it is also desirable to coupling reaction is carried out respectively when preparing different derived peptides, lacks that can to carry out Solid-phase synthesis peptides in batches anti-
The device and method answered.
The content of the invention
Solid-phase synthesis peptides can be carried out it is an object of the invention to provide a kind of by batch when preparing different derived peptides
The device of reaction.
To achieve the above object, multichannel Solid-phase synthesis peptides device of the invention includes the reaction for being used to contain reaction solution
Cylinder body, reaction cylinder base is connected with suction filtration adapter and nitrogen adapter, and nitrogen adapter is connected with nitrogen flexible pipe, and nitrogen flexible pipe is used for
External high-pressure nitrogen bottle is connected, nitrogen adapter is provided with nitrogen valve, and suction filtration adapter is provided with suction filtration valve；
Suction filtration adapter, which is connected with, vacuumizes flexible pipe, and vacuumizing flexible pipe is used to connect external Suction filtration device；Set at the top of reaction cylinder body
There is gas inlet/outlet pipe,
Reaction cylinder deck is connected with some connection grounds, and the top of connection ground is in up big and down small hollow taper type；It is each
A Peptide systhesis pipe, the bottom of the bottom intercalation reaction cylinder body of Peptide systhesis pipe and Peptide systhesis are inserted with respectively in connection ground
The bottom of pipe is provided with core filter core；The middle part of Peptide systhesis pipe is provided with frosted connecting portion, and frosted connecting portion is in being connected ground phase
The up big and down small hollow taper type of adaptation.
The bottom of each Peptide systhesis pipe is located in same level, reaction cylinder body to be set below each Peptide systhesis pipe
There is nitrogen point gas tank, top surface and each Peptide systhesis pipe of nitrogen point gas tank are corresponded provided with some nitrogen point gas port, nitrogen point
Check valve provided with one-way conduction from the bottom to top at gas port；The nitrogen adapter stretches into reaction cylinder body and divides gas tank phase with nitrogen
Connection.
The gas inlet/outlet pipe is provided with two, and one is connected at reaction cylinder body end face center, and another is connected to reaction
At the top of cylinder side wall.
The present invention devises a kind of multichannel Solid-phase synthesis peptides device, carries out Solid-phase synthesis peptides and reacts very convenient,
And batch reaction is carried out by the way that different Peptide systhesis pipes is inserted into same reaction cylinder body simultaneously, it is compared with the past to carry significantly
The high efficiency of Solid-phase synthesis peptides, has saved the time of corresponding research.Based on a kind of female peptide, various mutations position is being synthesized
,, can Fast back-projection algorithm be a variety of spreads out with prominent high efficiency advantage compared with conventional synthesizer during the different derived peptides of point
Raw peptide, so as to improve searching effect（Such as drug effect）The efficiency of optimal polypeptides matter.
In conventional Solid-phase Polypeptide reaction, resin and reaction solution add same Solid-phase Polypeptide reaction tube, and core filter core is then
Positioned at the middle part of many solid phase reaction pipes.This structure also limit the possibility of batch reaction.
Coupling Solution of the present invention when in use according to required for carrying out next step reaction is divided each Peptide systhesis pipe
Group, it would be desirable to Coupling Solution identical Peptide systhesis pipe be classified as same group so as to realize that batch reaction provides basis；This hair
The bright further technical schemes for setting out some Peptide systhesis pipes to correspond to same reaction cylinder bodies.
The method of this packet, and multitube correspond to the structure of same cylinder body, are that inventor is passing through long-term poorly efficient repetition
After reaction experiment, put forward by creative thinking, be to realize batch reaction, improve the key of reaction efficiency.
The gas inlet/outlet pipe is provided with two, and one is connected at reaction cylinder body end face center, and another is connected to reaction
At the top of cylinder side wall.Increasing the dynamics of nitrogen gas stirring（Nitrogen gas stirring dynamics can be increased by increasing nitrogen pressure）When, it can make
Nitrogen swimmingly flows out reaction cylinder body.
The setting of nitrogen point gas tank is another innovation of inventor, the bottom of each Peptide systhesis pipe（Core filter core）
Correspond to a nitrogen point gas port so that nitrogen drives part reaction cylinder body after by nitrogen point gas port, easily
Interior liquid enters Peptide systhesis pipe upwardly through core filter core, substantially increases stirring dynamics, hence it is evident that improves reaction or washes
The efficiency washed.
Brief description of the drawings
The multichannel Solid-phase synthesis peptides device that Fig. 1 is the present invention removes the structural representation after Peptide systhesis pipe；
Fig. 2 is Fig. 1 top view；
Fig. 3 is the schematic diagram that nitrogen point gas tank is connected with nitrogen adapter；
Fig. 4 is the structural representation of the multichannel Solid-phase synthesis peptides device of the present invention.
Embodiment
As shown in Figures 1 to 4, multichannel Solid-phase synthesis peptides device of the invention, including for containing reaction solution（Remove-insurance
Protect solution or Coupling Solution）Reaction cylinder body 1, reaction cylinder body 1 bottom is connected with suction filtration adapter 2 and nitrogen adapter 3, nitrogen adapter
3 are connected with nitrogen flexible pipe 4, and nitrogen flexible pipe 4 is used to connect external high-pressure nitrogen bottle, and nitrogen adapter 3 is provided with nitrogen valve 5, taken out
Filter adapter 2 and be provided with suction filtration valve 6；
Suction filtration adapter 2, which is connected with, vacuumizes flexible pipe 7, and vacuumizing flexible pipe 7 is used to connect external Suction filtration device；Reaction cylinder body 1 is pushed up
Portion is provided with gas inlet/outlet pipe 8,
The reaction top surface of cylinder body 1 is connected with some connection grounds 9, and the top of connection ground 9 is in up big and down small hollow taper type；Often
Be inserted with a Peptide systhesis pipe 10 respectively in one connection ground 9, the bottom of the bottom intercalation reaction cylinder body 1 of Peptide systhesis pipe 10 and
The bottom of Peptide systhesis pipe 10 is provided with core filter core 11；When being reacted, resin is placed on core filter core 11, and liquid can lead to
Core filter core 11 is crossed, but the resin of solid-state will not pass through core filter core 11.The middle part of Peptide systhesis pipe 10 is connected provided with frosted
Portion, frosted connecting portion is in being connected the up big and down small hollow taper type that ground 9 is adapted.In use, Peptide systhesis pipe 10 according to
It is supported on connection ground 9 and is sealed with being connected to be formed between ground 9 by frosted connecting portion.Frosted connecting portion is connected in Fig. 4
Connect ground 9 to be blocked, therefore do not provide label in figure.
The bottom of each Peptide systhesis pipe 10 is located in same level, reaction cylinder body 1 in each Peptide systhesis pipe 10
Lower section is corresponded provided with some nitrogen point provided with nitrogen point gas tank 12, top surface and each Peptide systhesis pipe 10 of nitrogen point gas tank 12
Check valve provided with one-way conduction from the bottom to top at gas port 13, nitrogen point gas port 13；The nitrogen adapter 3 stretches into reaction cylinder body 1
It is interior and be connected with nitrogen point gas tank 12.Check valve can prevent liquid from flowing into nitrogen point gas tank 12 by reaction cylinder body 1.Check valve
It is not shown for routine techniques.
The gas inlet/outlet pipe 8 is provided with two, and one is connected at the reaction end face center of cylinder body 1, and another is connected to instead
Answer the top side wall of cylinder body 1.When increasing the dynamics of nitrogen gas stirring, nitrogen can be made swimmingly to flow out reaction cylinder body 1.
Come according to the following steps when carrying out multichannel Solid-phase synthesis peptides using above-mentioned multichannel Solid-phase synthesis peptides device
Carry out：
First step is Connection Step, and nitrogen flexible pipe 4 is connected with external high-pressure nitrogen bottle, will vacuumize flexible pipe 7 with it is external
Suction filtration device be connected；
Second step is resin packing step, and essence is respectively charged into each Peptide systhesis pipe of multichannel Solid-phase synthesis peptides device
The resin for being connected with identical or different amino acid residue after really weighing；
Third step is to carry out resin swelling, is specifically：
1. swelling solvent is added into reaction cylinder body 1；
2. it will be equipped with each Peptide systhesis pipe of resin and insert same reaction cylinder body 1 by connecting ground 9 respectively being swelled, it is many
Peptide symthesis pipe is supported on connection ground 9 and its bottom is higher than nitrogen point gas tank 12 and positioned at the surface of nitrogen point gas port 13；
3. the air outlet valve and nitrogen valve 5 of high-pressure nitrogen bottle are opened, by nitrogen feeding nitrogen point gas tank 12；In nitrogen point gas tank 12
Nitrogen by after check valve enter reaction cylinder body 1, nitrogen drive part reaction cylinder body 1 in liquid entered by core filter core 11
Enter Peptide systhesis pipe, play a part of stirring in the reaction, liquid is fully contacted with resin；Nitrogen is closed by each polypeptide simultaneously
Flowed out into the gas inlet/outlet pipe 8 on the top end opening or reaction cylinder body 1 of pipe；It is swelled 30 ± 10 minutes, treats that resin is fully swelled
Afterwards, nitrogen valve 5 is closed；Due to the presence of check valve, the liquid in reaction cylinder body 1 will not enter nitrogen point gas by check valve
Case 12.
4. suction filtration operation is carried out, suction filtration operation is specifically to open suction filtration valve 6, and confining gas inlet/outlet pipe 8 starts suction filtration dress
Put（Such as vavuum pump）, suction filtration is carried out to reaction cylinder body 1, the liquid reacted in cylinder body 1 is extracted out out of reaction cylinder body 1；Suction filtration knot
Suction filtration valve 6 is closed after beam；
Four steps is to carry out deprotection reaction, is specifically：
1. deprotection solution is added into reaction cylinder body 1 by connecting ground 9 or Peptide systhesis pipe；
2. nitrogen valve 5 is opened, nitrogen gas stirring is carried out, the deprotection solution in reaction cylinder body 1 is fully contacted with resin；Nitrogen
Flowed out by the gas inlet/outlet pipe 8 on the top end opening or reaction cylinder body 1 of each Peptide systhesis pipe, this step persistently carries out 30-
40 minutes, nitrogen valve 5 is closed, the suction filtration operation is carried out；
5th step is washing resin steps, and this step is by solvent（Such as DMF solvent, methanol solvate or carbon dichloride solvent；
DMF solvent is DMF solvent）Add and resin is washed in reaction cylinder body, open nitrogen valve 5, carry out
Nitrogen gas stirring, makes the liquid in reaction cylinder body 1 fully be contacted with resin；Nitrogen by the top end opening of each Peptide systhesis pipe or
Gas inlet/outlet pipe 8 on reaction cylinder body 1 is flowed out, and this step is persistently carried out 1 minute, closes nitrogen valve 5, carries out the suction filtration behaviour
Make；
This step repeats six times；
6th step is the step of whether detection deprotection reaction completes, and is specifically：
Whether the deprotection reaction for detecting the resin in each Peptide systhesis pipe by ninhydrin completes, de- for not completing
The resin of reaction is protected, the 4th and the 5th step is repeated；
Resin for having completed deprotection reaction, if solid phase synthesis does not terminate all, carries out next step；If solid phase
Synthetic reaction all terminates, then takes out resin from each Peptide systhesis pipe, stops reaction；Here it is the knot of this synthetic method
Beam position.
7th step is to carry out coupling reaction, is specifically：
1. each Peptide systhesis pipe is made a distinction, the Coupling Solution according to required for carrying out next step reaction is to each Peptide systhesis pipe
Be grouped, it would be desirable to Coupling Solution identical Peptide systhesis pipe be classified as same group；（The Coupling Solution that next step reaction needs
Type is to need the amino acid being coupled to determine according to next step）
2. for the same Peptide systhesis pipe for being assembled with resin, by each Peptide systhesis pipe in the group respectively by connecting ground
The same reaction cylinder body 1 of 9 insertions, reaction cylinder body 1 is added by Coupling Solution；
3. nitrogen valve 5 is opened, nitrogen gas stirring is carried out, the liquid in reaction cylinder body 1 is fully contacted with resin；Nitrogen passes through each
Gas inlet/outlet pipe 8 on the top end opening or reaction cylinder body 1 of Peptide systhesis pipe is flowed out, and this step is persistently carried out 2-3 hours；Close
Nitrogen valve 5 is closed, the suction filtration operation is carried out；Then it is washing resin steps to carry out the 5th step, then carries out the 8th step；
8th step is the step of whether detection coupling reaction completes, and is specifically：
Whether the coupling reaction for detecting the resin in each Peptide systhesis pipe by ninhydrin completes；For not completing coupling
The resin of reaction, repeats the 7th step to re-start coupling reaction；Resin for having completed coupling reaction, then carry out the 4th
To the 6th step.
Wherein, the swelling solvent is DMF solvent；The resin uses Wang resins or Rink trees
Fat.
The present invention includes but is not limited to above-mentioned technical proposal, although having been carried out specifically to the present invention with reference to above-described embodiment
It is bright, it will be understood by those within the art that：The present invention can still be modified or equivalent substitution, without departing from
Any modification or partial replacement of the spirit and scope of the present invention, it all should cover among scope of the presently claimed invention.
Claims (3)
1. multichannel Solid-phase synthesis peptides device, it is characterised in that：Including the reaction cylinder body for containing reaction solution, cylinder body is reacted
Bottom is connected with suction filtration adapter and nitrogen adapter, and nitrogen adapter is connected with nitrogen flexible pipe, and nitrogen flexible pipe is used to connect external high pressure
Nitrogen cylinder, nitrogen adapter is provided with nitrogen valve, and suction filtration adapter is provided with suction filtration valve；
Suction filtration adapter, which is connected with, vacuumizes flexible pipe, and vacuumizing flexible pipe is used to connect external Suction filtration device；Set at the top of reaction cylinder body
There is gas inlet/outlet pipe,
Reaction cylinder deck is connected with some connection grounds, and the top of connection ground is in up big and down small hollow taper type；It is each
A Peptide systhesis pipe, the bottom of the bottom intercalation reaction cylinder body of Peptide systhesis pipe and Peptide systhesis are inserted with respectively in connection ground
The bottom of pipe is provided with core filter core；The middle part of Peptide systhesis pipe is provided with frosted connecting portion, and frosted connecting portion is in being connected ground phase
The up big and down small hollow taper type of adaptation.
2. multichannel Solid-phase synthesis peptides device, it is characterised in that：The bottom of each Peptide systhesis pipe is located at same level,
In the lower section of each Peptide systhesis pipe provided with nitrogen point gas tank, the top surface and each Peptide systhesis pipe one of nitrogen point gas tank in reaction cylinder body
One is correspondingly provided with the check valve provided with one-way conduction from the bottom to top at some nitrogen point gas port, nitrogen point gas port；The nitrogen connects
Pipe is stretched into reaction cylinder body and is connected with nitrogen point gas tank.
3. multichannel Solid-phase synthesis peptides device, it is characterised in that：The gas inlet/outlet pipe is provided with two, and one is connected to reaction
Cylinder deck center, another is connected at the top of reaction cylinder side wall.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
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| CN201710135871.8A CN107056879A (en) | 2017-03-08 | 2017-03-08 | Multichannel Solid-phase synthesis peptides device |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710135871.8A CN107056879A (en) | 2017-03-08 | 2017-03-08 | Multichannel Solid-phase synthesis peptides device |
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| CN107056879A true CN107056879A (en) | 2017-08-18 |
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| CN201710135871.8A Pending CN107056879A (en) | 2017-03-08 | 2017-03-08 | Multichannel Solid-phase synthesis peptides device |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2020103130A1 (en) * | 2018-11-23 | 2020-05-28 | 刘伟 | Microwave fluid solid-phase polypeptide synthesis method and system |
| US11319339B2 (en) | 2018-08-08 | 2022-05-03 | Hunan University Of Science And Engineering | Multi-channel peptide synthesizer and operating method thereof |
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| DE19602464A1 (en) * | 1996-01-24 | 1997-07-31 | Wolfgang Dr Rapp | Apparatus for multiple, simultaneous and parallel synthesis of chemicals |
| US5702672A (en) * | 1992-10-08 | 1997-12-30 | Warner-Lambert Company | Apparatus and method for multiple simultaneous synthesis |
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| CN203569010U (en) * | 2013-11-18 | 2014-04-30 | 苏州强耀生物科技有限公司 | Novel ultrasonic micro-shock polypeptide synthesizer |
| CN205662480U (en) * | 2016-06-08 | 2016-10-26 | 河南工程学院 | Polypeptide solid -phase synthesis pipe |
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2017
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1991013084A1 (en) * | 1990-02-22 | 1991-09-05 | Boehringer Ingelheim Kg | Process and device for the simultaneous synthesis of several polypeptides |
| US5702672A (en) * | 1992-10-08 | 1997-12-30 | Warner-Lambert Company | Apparatus and method for multiple simultaneous synthesis |
| DE19602464A1 (en) * | 1996-01-24 | 1997-07-31 | Wolfgang Dr Rapp | Apparatus for multiple, simultaneous and parallel synthesis of chemicals |
| WO1998006490A1 (en) * | 1996-08-12 | 1998-02-19 | Monsanto Company | Solid phase organic synthesis device with pressure-regulated manifold |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US11319339B2 (en) | 2018-08-08 | 2022-05-03 | Hunan University Of Science And Engineering | Multi-channel peptide synthesizer and operating method thereof |
| WO2020103130A1 (en) * | 2018-11-23 | 2020-05-28 | 刘伟 | Microwave fluid solid-phase polypeptide synthesis method and system |
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