JPH0642937U - Peptide fragment fractionation device - Google Patents

Abstract

(57) [Summary] [Purpose] To reduce the amount of reagents remaining in the piping. A peptide fragment fractionation device is a device for fractionating a carboxyl-terminal peptide from a peptide fragment mixture, and comprises a reaction container, a coupling means, a reagent supply means, a collection means, and a reagent collection means. Here, the reagent recovery means recovers the reagent remaining in the pipe into the reagent storage section.

Description

[Detailed description of the device]

[0001]

[Industrial applications]

 The present invention is directed to a peptide fragment fractionation device, particularly to extract a carboxyl-terminal peptide from a peptide fragment mixture in which the peptide bond between the lysine residue in the polypeptide and the subsequent amino acid residue on the carboxyl-terminal side is specifically cleaved. The present invention relates to a peptide fragment fractionation device for fractionation.

[0002]

[Prior art and its problems]

 As a method for analyzing the carboxyl terminus (C terminus) of a peptide, there is a peptide fractionation method disclosed in JP-A-1-235600. In this peptide fractionation method, the peptide bond between the lysine residue in the polypeptide and the subsequent C-terminal amino acid residue is specifically cleaved. Then, the obtained peptide fragment mixture is reacted with a solid support having a functional group capable of reacting with a free amino group to form a covalent bond. Then, the peptide bond between the terminal residue having the amino group at the α-position of each peptide and the residue adjacent to it is cleaved with trifluoroacetic acid (TFA), and the C-terminal peptide fragment of the released protein is collected. To do.

As an apparatus for carrying out this peptide fractionation method, a reaction container filled with a peptide fragment mixture and a solid phase support, and a coupling for covalently binding each peptide of the peptide fragment mixture and the solid phase support And a reagent supply unit for supplying a reagent containing an acid (particularly TFA is suitable) from the reagent bottle to the reaction container via a pipe, and a recovery unit for recovering the cleaved fragments. The inventor thought. However, in the peptide fragment fractionation device having such a configuration, after the reagent such as TFA is supplied to the reaction container, the reagent remains in the pipe between the reagent bottle storing the reagent and the reaction container. . Therefore, the following problems occur.

[0004] As time passes, the TFA is partially decomposed and deteriorated, and the TFA of which the purity is lowered in the next liquid transfer affects the device performance and the reactivity itself. Since TFA penetrates the pipe and diffuses in the device, it causes corrosion of the device. An object of the present invention is to reduce the reagent remaining in the pipe.

[0005]

[Means for Solving the Problems]

 The peptide fragment fractionation device according to the present invention is capable of fragmenting a carboxyl-terminal peptide fragment from a peptide fragment mixture in which a peptide bond between a lysine residue in a polypeptide and a subsequent amino acid residue on the carboxyl terminal side is specifically cleaved. It is a device for sorting. This device comprises a reaction container, a coupling means, a reagent supply means, a recovery means, and a reagent recovery means.

The reaction vessel is filled with a solid phase support and the peptide fragment mixture is injected. The coupling means adjusts the temperature in the reaction vessel to covalently bond the peptide fragment mixture to the solid support. The reagent supply means has a reagent storage part for storing the reagent, and supplies the reagent from the reagent storage part to the reaction container through a pipe. The recovery means is for recovering the peptide fragment from the reaction vessel. The reagent recovery means collects the reagent remaining in the pipe in the reagent storage section.

[0007]

[Action]

 In the peptide fragment fractionation device according to the present invention, the reaction container is filled with the solid phase support, and when the peptide fragment mixture is injected therein, the temperature in the reaction container is adjusted by the coupling means, The peptide fragment mixture is coupled to a solid support. Then, the reagent supply means supplies the reagent from the reagent storage section to the reaction container through the pipe. Further, the peptide fragment from the reaction container is recovered by the recovery means. When these processes are completed, the reagent remaining in the pipe is recovered in the reagent storage section by the reagent recovery means.

[0008] Here, the reagent recovery means ensures that the reagent remaining in the pipe is collected in the reagent storage section. Therefore, the reagent remaining in the pipe is reduced. This improves the life of the device and reduces the risk of contamination on next use.

[0009]

【Example】

 A peptide fragment sorting apparatus 1 according to an embodiment of the present invention shown in FIG. 1 has a box-shaped main body case 2 and an operation panel 3 arranged on the front upper part of the main body case 2. A first container placement part 4 and a second container placement part 5 are arranged on the front side of the main body case 2 in an upper and lower direction.

In the first container arranging section 4, a recovery bottle attaching section 7 for attaching a recovery bottle 6 for accommodating the recovered C-terminal peptide and a heat block 8 accommodating a column 32 described later are arranged side by side. Has been done. A waste liquid bottle mounting portion 10 for mounting a waste liquid bottle 9, a recovery liquid bottle mounting portion 12 for mounting a recovery liquid bottle 11 for storing a recovery liquid, and trifluoroacetic acid used for cutting are disposed in the second container placement portion 5. A cutting bottle mounting portion 14 for mounting the cutting liquid bottle 13 storing (TFA) and a cleaning bottle mounting portion 15 for mounting the cleaning liquid bottle 15 storing the cleaning liquid are juxtaposed.

As shown in FIG. 2, the operation panel 3 includes a process display section 19 in which eight LEDs 1 to LED 8 are arranged in parallel, and a key input section 20 in which five keys K1 to K5 are arranged in parallel. , And a status display section 21 in which six LEDs are vertically arranged. LED1 is used in the coupling process, LED2 is used in the first cleaning process, LED3 is used in the second cleaning process, LED4 is used in the drying process, LED5 is used in the cutting process, LED6 is used in the collecting process, LED7 is used in the concentrating process, and LED8 is used. Lights up when the process is completed.

The key K1 is a washing key (WASH) which is operated only when washing the inside of the pipe. The key K2 is a TFA recovery key (TFA / RET) operated at the time of TFA recovery described later. The key K3 is a cursor key for making progress from an arbitrary process. The key K4 is a run key (RUN) for automatically advancing a process. The key K5 is a stop key (STOP) for forcibly stopping the process.

In the main body case 1, a piping system including 13 solenoid valves SV1 to SV13 shown in FIG. 3 is arranged. The solenoid valves SV1 to SV7 and SV11 to SV13 are normally closed two-way valves. These solenoid valves open when energized. The solenoid valves SV8 to SV10 are three-way valves, of which two inlet ports are normally open and the remaining outlet ports are normally closed. Therefore, the solenoid valves SV8 to SV10 open all ports when energized and close only the outlet port when shut off.

A nitrogen gas cylinder (not shown) is connected to the inlet port of the solenoid valve SV1. The inlet ports of the solenoid valves SV2 to SV7 are collectively connected to the outlet port of the solenoid valve SV1. The outlet port of the solenoid valve SV2 is connected to the column 32 in the heat block 8 via the solenoid valve SV8, the solenoid valve SV9, the measuring pipe 31, and the solenoid valve SV10.

A proximal end of a pipe whose tip is inserted into the cleaning bottle 15 is connected to the outlet port of the solenoid valve SV3. The proximal end of the pipe whose tip is inserted into the recovery liquid bottle 11 is connected to the outlet port of the solenoid valve SV4. The base end of the pipe whose front end is inserted into the cutting liquid bottle 13 is connected to the outlet port of the solenoid valve SV5. The electromagnetic valve SV6 is connected to the base end of a pipe whose front end is inserted into the waste liquid bottle 9. The proximal end of the pipe whose tip is inserted into the recovery bottle 6 is connected to the outlet port of the solenoid valve SV7.

The solenoid valve SV8 has an outlet port connected to the proximal end of a pipe whose tip is inserted near the bottom surface of the cleaning bottle 15. The base end of a pipe whose tip is inserted near the bottom surface of the recovery liquid bottle 11 is connected to the outlet port of the solenoid valve SV9. The outlet port of the solenoid valve SV10 is connected to the proximal end of a pipe whose tip is inserted near the bottom surface of the cutting liquid bottle 13. Note that these pipes are made of transparent resin tubes in consideration of corrosion resistance and the like.

In the heat block 8, a column 32 for packing the enzyme-treated peptide and the solid phase support can be arranged. A distribution block 34 is connected to the column 32 via a pipe. A liquid sensor 33 is arranged between the column 32 and the distribution block 34 to detect the liquid passing through the pipe to detect that the column 32 is filled with the liquid. The distribution block 34 is branched in two directions. One side is connected with the pipe inserted into the waste bottle 9, and the other side is connected with the pipe inserted into the recovery bottle 6. Further, the cutting liquid bottle 13, the waste liquid bottle 9 and the recovery bottle 6 are connected to the inlet ports of the solenoid valves SV11, SV12 and SV13 via pipes. The outlet ports of these solenoid valves SV11, SV12, SV13 are open to the atmosphere.

The peptide fragment sorting apparatus 1 has a control unit 40 shown in FIG. The control unit 40 is composed of a microcomputer including memories such as ROM and RAM. Solenoid valves SV1 to SV13 are connected to the control unit 40. The control unit 40 has keys K1 to K5, process and status display units 19 and 21, a temperature control unit 41 for controlling the temperature of the heat block 8, and a processing time for each process. The timer 42 and the liquid sensor 33 are connected. The temperature control unit 41 is provided with a temperature setting unit (not shown).

Next, the control operation of the peptide fragment sorting apparatus 1 configured as described above will be described with reference to the flowcharts shown in FIGS. Before starting a specific control operation, the operator fills each bottle with a reagent and attaches it to each attachment part. Here, the cleaning bottle 9 is filled with methanol or acetnitrile. The recovery liquid bottle 11 is filled with a 50% acetonitrile / 2-propanol mixed liquid containing 0.1% TFA. The cutting liquid bottle 13 is filled with TFA.

Further, the heat block 8 is equipped with a column 32 filled with a solid-phase support having a functional group (porous glass having an isociothiate group introduced). Column 32 is pre-injected with a peptide fragment mixture containing amino acid residues treated with the enzyme (lysyl endopeptidase). The urea used in the enzyme treatment remains in this peptide fragment mixture.

On the other hand, in the control unit 40 of the peptide fragment fractionation device 1, when the program starts, first, in step S1, initialization is performed. Here, for example, the memory in the control unit 40 is cleared. In step S2, the variable s indicating the progress of the process is set to "1". In step S3, the LED (first LED1) of the process display unit 19 is turned on.

Then, the key determination process is repeated in steps S4 to S7. Here, in step S4, it is determined whether or not the run key K3 has been operated. In step S5, it is determined whether or not the cleaning key K1 has been operated. In step S6, it is determined whether the TFA recovery key K2 has been operated. In step S7, it is determined whether or not the cursor key K3 has been operated.

If it is determined in step S4 that the run key K3 has been operated, the process proceeds to step S8. In step S8, the sequential process (RUN process) shown in FIG. 7 is executed. If it is determined in step S5 that the cleaning key K1 has been operated, the process proceeds to step S9. In step S9, a cleaning process for cleaning the inside of the pipe is executed. If it is determined in step S6 that the TFA recovery key K2 has been operated, the process proceeds to step S10. In step S10, the TFA recovery process shown in FIG. 16 is executed. If it is determined in step S7 that the cursor key K3 has been operated, the process proceeds to step S11. In step S11, cursor processing is executed. In this cursor processing, the variable s is incremented each time the cursor key K3 is operated. The variable s is changed so that the value after "7" returns to "1". Also, when the variable s is "8", it is changed so as to return to "1".

In the sequential processing in step S8, the variable s is first read in step S31 in FIG. In step S32, it is determined whether or not the value of the read variable s is "1". When it is determined that the variable s is "1", the process proceeds to step S33. In step S33, a coupling treatment between the solid phase support packed in the column 32 and the peptide fragment mixture is performed. In step S35, the unbound peptide is purged and the first cleaning with the recovery liquid is performed. This flushes the unbound peptide fragment mixture. In step S35, the second cleaning with the cleaning liquid is performed. This cleans the column 32 and the inside of the pipe. In step S37, a drying process with N ₂ gas is performed. In step S38, a cutting process is performed by acid treatment using TFA. In step S39, a fragment collection process using a collection liquid is performed. In step S40, the collected fragment is concentrated with N ₂ gas.

When these processes are completed, the variable s is set to “8” in step S41 so that the completion of the series of separation processes can be notified by turning on the LED 8. Then return to the main routine. When it is determined in step S32 that the variable s is not "1" (when the process is started from an intermediate step), the variable s is set to any value of "2" to "6" in steps S43 to S47. Determine if there is. Then, depending on the determination result, the process shifts from any of steps S43 to S47 to any of steps S36 to S39. When the variable s is "7", the process moves from step S47 to step S40.

In the coupling process of step S33, the LED of the RUN key is turned on in step S51 of FIG. This allows the operator to recognize that a series of processes has started. In step S53, the temperature of the heat block 8 is controlled by the temperature controller 41 within the range of 4 ° C to 80 ° C (preferably 10 ° C to 60 ° C). This temperature control is performed for a set time T ₂ (for example, 5 minutes to 3 hours). That is, waits for the elapse of the set time T ₂ at step S54, and ends the temperature control proceeds to step S55 after a lapse of the set time T _2. In step S56, LED1 is turned off. Thereby, the coupling reaction between the solid phase support and the peptide fragment mixture is carried out.

In the first cleaning step of step S35 of FIG. 7, the LED 2 is turned on in step S60 of FIG. In step S61, the solenoid valves SV1, 7, 12 are opened. Then, the inside of the recovery bottle 6 is pressurized with N ₂ gas, and the waste liquid bottle 9 is opened to the atmosphere. Then, N ₂ gas is supplied to the column 32 via the solenoid valves SV1, _2, 8, 9, 10 and thereby the uncoupled peptide in the column 32 is purged into the waste liquid bottle 9. . In the step S62 waits for the lapse of the set time T _3. If it is determined that the set time has elapsed, the process proceeds to step S63. In step S63, the solenoid valves SV1, 2, 7, 12 are closed. In step S72 of FIG. 10, the solenoid valves SV1, 4, 9, 7, 12 are opened. As a result, the N ₂ gas is filled in the recovery liquid bottle 11 and also in the recovery bottle 6. As a result, the pressure inside the recovery liquid bottle 11 and the recovery bottle 6 rises above atmospheric pressure, and the recovery liquid filled in the recovery liquid bottle 11 is supplied to the column 32 via the solenoid valves SV9, 10. In step S73, the liquid sensor 33 waits until the column 32 is filled with the recovery liquid.

When the liquid sensor 33 detects the flow of the recovery liquid and the column 32 is filled with the recovery liquid, the process proceeds to step S74. In step S74, the solenoid valves SV1, 4, 9, 7, 12 are closed. Then, in step S75, the solenoid valves SV1, SV2, SV7, SV12 are opened in order to discharge the recovery liquid in the column 32 and the pipe. As a result, the recovery liquid bottle 6 is pressurized and the waste liquid bottle 9 is opened to the waste liquid bottle 9 side via the distribution block 34. Emitted. In step S76, the elapse of the set time T ₄ is awaited. When the set time T _{4 has} elapsed, the process proceeds to step S77. In step S77, the solenoid valves SV1, 7, 12 are closed. Then, in step S78, the LED 2 is turned off to notify that the first cleaning process is completed, and then the process proceeds to step S36 in FIG.

In the second cleaning step of step S36 of FIG. 7, the LED 3 is turned on in step S81 of FIG. Then, in step S82, the solenoid valves SV1, 3, 8, 7, 12 are opened. As a result, the cleaning bottle 11 and the recovery liquid bottle 6 are filled with N ₂ gas and the inside is pressurized. Further, the inside of the waste liquid bottle 9 is opened to the atmosphere. Then, the cleaning liquid stored in the cleaning bottle 15 is supplied into the column 32 via the solenoid valves SV8, 9, 10. The cleaning liquid supplied into the column 32 flows to the waste liquid bottle 9 side as in the first cleaning. In step S83, the liquid sensor 33 waits for the cleaning liquid to fill the column 32. When the liquid sensor 33 detects the cleaning liquid, the process proceeds to step S84. In step S84, the opened solenoid valves 1, 3, 8, 7, 12 are closed. The subsequent operation in steps S85 to S88 is the same as the operation in steps S75 to S78 in the first cleaning, and thus the description thereof is omitted.

In the drying process of step S37 of FIG. 7, the LED 4 is turned on in step S91 of FIG. At step S92, the solenoid valves SV1, SV2, SV7, SV12 are opened and N ₂ gas is supplied into the column 32. In step S93, the elapse of the predetermined time T ₆ is awaited. When the set time T ₆ has elapsed, the process proceeds to step S94. When the set time T ₆ has elapsed, the process proceeds to step S94, and the opened solenoid valves SV1, SV2, S7, S12 are closed. Then, the process proceeds to step S95, the LED 4 is turned off, and the process proceeds to step S38 in FIG. In addition, here, the drying process can be simplified because it is dried with N ₂ gas instead of being dried in a vacuum.

In the cutting step of step S38 of FIG. 7, the LED 5 is turned on in step S101 of FIG. In step S102, the solenoid valves SV1, 5, 10, 7, 12 are opened respectively. As a result, the TFA stored in the cutting liquid bottle 13 is supplied into the column 32 via the solenoid valve SV10. In step S103, it waits for the liquid sensor 33 to detect the TFA liquid. When it is determined that the column 32 is filled with TFA and the liquid sensor 33 detects the TFA liquid, the process proceeds to step S104. In step S104, the solenoid valves SV1, 5, 10, 7, 12 are closed.

In step S 105, the temperature of the heat block 8 is adjusted by the temperature adjusting unit 41. The temperature at this time is in the range of 20 ° C to 80 ° C (preferably 30 ° C to 60 ° C). In step S106, the setting time T ₇ (5 minutes to 1 hour) elapses. When set time T ₇ in step S1 06 is determined to have elapsed the process proceeds to step S107. In step S107, the temperature control ends. In step S108, the solenoid valves SV1, SV2, SV6, SV13 are opened. As a result, the inside of the waste liquid bottle 9 is pressurized and the inside of the recovery bottle 6 is opened to the atmosphere. Then, N ₂ gas is ejected toward the collection bottle 6 side through the column 32 and the distribution block 34, so that the inside of the column 32 is purged. In step S109, the setting time T ₈ is awaited. When the set time T ₈ has elapsed, the process proceeds to step S110, and the solenoid valves SV1, 2, 6, 13 are closed. Then, after turning off the LED 5 in step S111, the process proceeds to step S39 in FIG.

In the recovery process of step S39 of FIG. 7, the LED 6 is turned on in step S121 of FIG. In step S122, the solenoid valves SV1, 4, 9, 6, 13 are opened. As a result, the inside of the recovery liquid bottle 11 and the waste liquid bottle 9 is pressurized, and the inside of the recovery bottle 6 is opened to the atmosphere. Accordingly, the recovery liquid in the recovery liquid bottle 11 is supplied into the column 32 via the electromagnetic valve SV9, the measuring pipe 31, and the electromagnetic valve SV10. In step S123, the liquid sensor 33 waits for detection of the collected liquid. When the liquid sensor 33 detects the recovered liquid, the process proceeds to step S124. In step S124, the solenoid valves SV1, 4, 9, 6, 13 are closed. Subsequently, in step S122, the solenoid valves SV 1, 2, 6, 13 are opened. As a result, the recovery liquid of the C-terminal peptide cleaved in the column 32 is recovered in the recovery bottle 6 via the distribution block 34. In step S126, the elapse of the set time T ₉ is awaited. When the set time T ₉ has elapsed, the process proceeds to step S127, and the solenoid valves SV1, SV2, S6, S13 are closed. As a result, the C-terminal peptide remaining in the column 32 is efficiently collected in the collection bottle 6, and the LED 6 is turned off in step S128, and then the process proceeds to step S40 in FIG.

In the concentration processing routine of step S40 of FIG. 7, the LED 7 is turned on in step S131 of FIG. In step S132, the solenoid valves SV1, 7, 13 are opened. As a result, N ₂ gas flows into the recovery bottle 6, and the recovery liquid in the recovery bottle 6 is evaporated and concentrated. In step S133, the elapse of the set time T ₁₀ is awaited. This set time is, for example, about 3 hours. At step S134, the solenoid valves SV1, 7, 13 are closed. Then, after turning off the LED 7 in step S135, the process proceeds to step S41 in FIG.

In step S41, the variable s is set to “1” in order to turn on the LED 8 in step S3 (FIG. 5). At the time when this series of sequential processing is completed, the inside of the cutting liquid bottle 13 is still pressurized, and TFA remains in the pipe connecting the solenoid valve SV10 and the cutting liquid bottle 13. To remove the TFA in this pipe, operate the TFA recovery key K2.

In the TFA recovery process of step S14 of FIG. 5, the LED provided on the TFA recovery key K2 is turned on in step S141 of FIG. At step S142, the solenoid valves SV1, SV2, S10, S11 are opened. As a result, the inside of the cutting liquid bottle 13 is opened to the atmospheric pressure, N ₂ gas flows to the cutting liquid bottle 13 through the solenoid valve SV10, and the TFA liquid remaining in the pipe is collected in the cutting liquid bottle 13. To be done. In step S143, the elapse of the set time T ₁₁ is awaited. The set time T ₁₁ is a time sufficient to collect the TFA liquid in the pipe. If it is determined that the set time T ₁₁ has elapsed, the process proceeds to step S144.

In step S144, the solenoid valves SV1, SV2, S10 are closed. In step S145, the elapse of the set time T ₁₂ is waited for. The set time T ₁₂ is sufficient to bring the cutting liquid bottle 13 to atmospheric pressure. When the set time T ₁₂ has elapsed, the process proceeds to step S146. Then, after closing the solenoid valve SV11 in step S146, the process returns to the main routine.

When it is desired to interrupt the processing in the middle of the series of sequential processing, the stop key K5 is pressed. When the stop key K5 is operated, interrupt processing is performed, and the stop processing shown in FIG. 6 is executed. In step S21 of FIG. 6, the operation during the sequential processing is temporarily stopped. In step S22, it is determined whether or not the stop key K5 has been operated again. If it is determined that the stop key K5 is not operated, the process proceeds to step S24. In step S24, it is determined whether or not the run key K4 has been operated. If the run key K4 is not operated, the process returns to step S21 and the suspended state is continued.

If the stop key K5 is operated again, the program proceeds from step S22 to step S23. In step S23, in order to return the process step to the beginning, the program counter is changed so that the next process starts from step S2 in FIG. 5, and this process is ended. If the run key K4 is operated, the process proceeds to step S25. In step S25, the suspended operation is restarted.

In the above-described embodiment, the column 32 is filled with the solid phase support, the peptide fragment mixture is injected, the block is mounted on the heat block 8, and each bottle is mounted on the corresponding mounting portion by a simple operation. , C-terminal fragments can be separated from a mixture of peptide fragments. Therefore, a C-terminal fragment can be easily and reproducibly separated from a peptide without requiring skill.

Further, since the reagent remaining in the pipe can be recovered by the TFA recovery process, diffusion of TFA vapor from the pipe can be reduced and troubles such as corrosion can be reduced. Furthermore, since the liquid from the column 32 is distributed by selectively opening the waste liquid bottle 9 and the recovery bottle 6 to the pressure and the atmospheric pressure, it is not necessary to provide a solenoid valve in the distribution flow path. For this reason, the dead volume on the downstream side of the reaction column or the like can be reduced, and long-term use is possible even if a sample or reagent that precipitates when dried, such as urea, is used.

[Other Embodiments] (a) In the above embodiment, the TFA recovery process is configured to be performed independently, but it may be automatically performed every time the cutting with the TFA solution is completed. (B) In the above embodiment, only the TFA for cleavage was collected, but other reagents may be collected.

[0043]

[Effect of device]

 In the peptide fragment fractionation device according to the present invention, the reagent remaining in the pipe is collected in the reagent storage portion by the reagent collecting means, so that the reagent remaining in the pipe is reduced. This greatly reduces the risk of contamination and also prolongs the life of the device significantly.

[Brief description of drawings]

FIG. 1 is a perspective external view of a peptide fragment sorting apparatus according to an embodiment of the present invention.

FIG. 2 is a front view of the operation panel.

FIG. 3 is a schematic view of the piping system.

FIG. 4 is a block diagram of its control system.

FIG. 5 is a control flowchart thereof.

FIG. 6 is a control flowchart thereof.

FIG. 7 is a control flowchart thereof.

FIG. 8 is a control flowchart thereof.

FIG. 9 is a control flowchart thereof.

FIG. 10 is a control flowchart thereof.

FIG. 11 is a control flowchart thereof.

FIG. 12 is a control flowchart thereof.

FIG. 13 is a control flowchart thereof.

FIG. 14 is a control flowchart thereof.

FIG. 15 is a control flowchart thereof.

FIG. 16 is a control flowchart thereof.

[Explanation of symbols]

 1 Peptide Fragment Separation Device 6 Recovery Bottle 8 Heat Block 13 Cutting Liquid Bottle 32 Reaction Vessel 40 Control Section SV5, SV11 Solenoid Valve

Claims (1)

[Scope of utility model registration request] 1. An apparatus for fractionating a carboxyl-terminal peptide fragment from a peptide fragment mixture in which a peptide bond between a lysine residue in a polypeptide and a subsequent amino acid residue on the carboxyl terminal side is specifically cleaved. A reaction container filled with the peptide fragment mixture and a solid support, a coupling means for covalently bonding the peptide fragment mixture to the solid support by adjusting the temperature in the reaction container, and a reagent container A reagent supply unit for supplying the reagent to the reaction container from the reagent storage unit via a pipe, a recovery unit for recovering a peptide fragment from the reaction container, and the pipe. And a reagent recovery means for recovering the remaining reagent in the reagent storage part. .