CEM Razor Installation And Operation Instructions Manual

Razor
Parallel Cleavage System
Installation and Operation Instructions
600850
Rev. 2
July 2017
Introduction to the Razor
The RazorTM parallel cleavage system is an isolated heating module designed for performing the peptide cleavage step. The isolated module provides the advantage of protecting the Liberty Blue system from the harsh reagents required for the cleavage all while allowing the use of heat to provide high quality peptides in excellent yields in a fraction of the time.
Razor Hardware
Vessel
Hot block
Valve Panel
Vessel Tray
Temperature
Control Panel
Heat
Switch
Locking Lever
Power
Switches
Vacuum
Switch
Vacuum
Gauge
Vent,
Vacuum,
and Waste
Ports
Razor™ Parallel Peptide Cleavage System
1
Hardware Installation
Installation Site Preparation
The Razor parallel cleavage system should be installed in a functioning laboratory fume hood. Choose a location that:
can accomodate the Razor system [15” x 11.5” x 15.5” (38.1 cm x 29.2 cm x 39.4 cm) (L x W x H)] as well as an
additional 8” (20 cm) of open space on each side and 6” (15 cm) at the rear of the instrument for proper ventilation.
is free from vibration of large equipment and/or excessive walk-through trac,
provides a temperature range of 41 - 104 ºF (5 - 40 ºC) and a humidity range of 10 - 85% relative humidity
provides adequate space for sample handling, and
permits the system to be connected to a dedicated, grounded 120 VAC outlet. The Razor should operated on a
stabilized, constant voltage AC power supply. To operate properly, the voltage must be within ±10% of the specied level.
Razor Hardware
The Razor accessory kit comes with the following items:
1. Razor vessel rack (P/N: 547205)
2. Razor vessel rack cover (P/N:
170875)
3. 1 L glass bottle (P/N: 167550)
4. Waste tubing bundle (P/N: 547225)
5. GL45 waste cap (P/N: 167545)
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6. Razor power supply (P/N: 267080)
7. Power cord
8. Razor vessel caps (P/N: 170875)
9. 50 mL centrifuge tubes, set of 25 (P/N: 330090)
10. Razor cleavage vessel, 120 pack (P/N: NP-1392) (not pictured)
Razor™ Parallel Peptide Cleavage System
Hardware Assembly and Setup
WARNING
The Razor should be installed in a ventilated fume hood.
1. Remove the Razor from it’s shipping box and place the system in a well ventilated fume hood.
2. Connect the power cord to the power supply box. Attach the power supply to the connector on the rear of the system, and plug the power cord into an appropriate outlet.
3. Attach the two tubes on one end of the waste tubing bundle to the GL45 cap. It does not matter which tube is connected to which hole. Secure the cap to the 1 L bottle.
4. Connect the labeled waste cap assembly tubes to the vacuum and vent ports on the side of the system.
Razor™ Parallel Peptide Cleavage System
3
5. Load the vessel tray with 12 clean 50 mL centrifuge tubes (P/N: 330090), and slide into the Razor cavity. Make sure the tray has been pushed all the way to the back of the cavity. Lock the vessel tray in place by pulling the locking lever down.
6. Switch the system power and cavity lights ON using the power switches on the rear of the system.
7. Switch the system heat ON using the power switch on the left side of the system.
8. Set and verify the system temperature.
8.1. Using the up and down arrows on the temperature screen, set the temperature to 40 - 42 ºC. The set
temperature is displayed in green in the lower right corner of the screen.
8.2. The current temperature is displayed in white in the center of the screen. The system is ready for use when
the current temperature matches the set temperature.
Current Temperature
Set Temperature
Change Temperature
Recommended Cleavage Parameters
In Fmoc synthesis, the removal of the peptide from the solid support is typically accomplished with triuoroacetic acid (TFA). Because the side chain protecting groups used in Fmoc synthesis are acid labile, a single step both cleaves the peptide from the resin and removes the protecting groups. Various scavenger molecules are added to the TFA to prevent the cleaved protecting groups from reattaching to the peptide. The particular scavengers used depend on the specic peptide sequence. Common scavengers include water (scavenges t-butyl cations), triisopropyl silane (TIS, scavenges trityl and Pbf cations), ethane dithiol (EDT, scavenges t-butyl cations, reduces oxidation of Cys/Met side chains), dioxa-1,8-octane-dithiol (DODT, scavenges t-butyl cations, suppresses oxidation of Cys/Met side chains), phenol (protects Tyr and Trp side chains from oxidation), and thioanisole (aids in removal of Pbf protecting groups from Arg(Pbf), suppresses oxidation of Cys/Met side chains).
CEM recommends using a cocktail of TFA/TIS/H2O/DODT (92.5/2.5/2.5/2.5).
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Razor™ Parallel Peptide Cleavage System
General Procedures
Razor Cleavage Protocol
1. Verify the current system temperature has reached the set temperature.
2. Load the vessel tray with 12 clean 50 mL centrifuge tubes (P/N: 330090), and slide into the Razor cavity. Make sure the tray has been pushed all the way to the back of the cavity. Lock the vessel tray in place by pulling the locking lever down.
CAUTION
The vessel tray must have all 12 centrifuge tubes loaded to operate correctly. If a tube is missing, the system will fail to create a vacuum during draining.
3. Insert a new, clean reaction tube into the washing station. Switch the vacuum pump ON using the power switch on the right side of the system. Verify there are no leaks by checking the vacuum gauge below the vacuum switch, and then turn the pump OFF.
NOTE
Up to 11 additional vessels can be washed and added to the Razor hotblock. To prepare an additional vessel, wash the resin as instructed while keeping the valves for the already prepared vessels in the CLOSED position to prevent premature draining.
An optional Teon® cap (P/N: 170810) can be placed on reaction vessels during cleavage to reduce evaporation.
4. Transfer the resin with the completed peptide into the reaction tube. Move the Drain valve knob to the OPEN position and turn the vacuum pump ON to drain any solvent from the resin.
OpenClosed
5. Rinse the resin three times with an appropriate volume of dichloromethane (DCM), leaving the pump ON and the Drain knob OPEN to drain the liquid. After rinsing, turn the vacuum pump OFF and CLOSE the Drain knob.
CAUTION
DMF is a base and will react with TFA. DCM should be used to rinse the resin prior to cleavage to ensure that no residual DMF is present before adding the cleavage cocktail.
Razor™ Parallel Peptide Cleavage System
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6. Move the reaction tube containing the rinsed resin to an empty, clean position in the Razor hot block and turn the numbered valve knob to the CLOSED position.
7. Add cleavage cocktail to the reaction vessel, double that of the volume of the resin bed. For 0.25 mmol cleavages, the resin mixture may be visible above the hot block. Up to 12 reaction vessels can be loaded into the hot block.
8. Allow the vessel(s) to remain in the Razor for 30 minutes each at 40 - 42 ºC.
NOTE
If the peptide has multiple Arg residues, increase the cleavage time by 5 min for every additional Arg after the third Arg residue. Do not exceed 45 min for the total cleavage time.
9. After cleavage has ended, turn the appropriate numbered valve knob to the OPEN position to begin draining the cleavage solution.
CAUTION
Drain one cleavage position at a time, do not drain multiple positions at once.
10. Turn the vacuum pump ON to drain any remaining liquid, then turn OFF.
11. Move the Vent valve knob to the OPEN position to release pressure.
12. Unlock the vessel tray by pushing the locking lever up. This will lower the tray for removal.
13. Carefully remove the vessel tray from the Razor cavity and remove the centrifuge tubes containing cleaved peptide. If moving the tray to a new location, the vessel tray cover (P/N: 170875) can be installed.
14. Replace the used 50 mL centrifuge vessels with clean vessels and move the Vent valve to the CLOSED position.
15. Run a cleaning procedure on each cleavage position used (see Maintenance Procedures).
16. To empty the waste bottle, remove the waste cap and discard the waste then rmly re-secure the cap.
Precipitation and Analysis
1. To precipitate the cleaved peptide, add ice cold ether up to 50 mL (or 8 times the amount of drained liquid). If the volume exceeds 50 mL, use a second centrifuge tube. The ether should be ice cold to ensure the maximum amount of peptide precipitates out.
2. Centrifuge the peptide solution for 5 min at 3500 rpm or until a white or clear peptide pellet forms on the bottom of the tube. Repeat the centrifuge process if necessary to remove any oating particles.
3. Decant the ether, leaving the precipitated peptide in the tube.
4. The peptide can be resuspended in ice cold ether and centrifuged again to ensure all protecting groups are gone.
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Razor™ Parallel Peptide Cleavage System
5. The peptide can be analyzed immediately on an HPLC or UPLC for purity, or can be lyophilized overnight.
6. To lyophilize, suspend the peptide gel in 1% acetic acid in deionized water then freeze with liquid nitrogen before placing on the lyophilizer.
Maintenance Procedures
Cleaning
Each vessel position should be cleaned before and after use to prevent cross contamination. To clean a vessel position:
1. Insert a 50 mL centrifuge tube in the vessel tray and lock the tray in position.
2. Insert an empty reaction vessel into the appropriate cleavage position.
3. Turn the vacuum pump ON and rinse ethanol or isopropanol through the reaction vessel.
WARNING
Do not use DCM to rinse any cleavage positions post-cleavage as this can lead to more frequent seal replacements.
4. Turn the vacuum pump OFF and vent the system by turning the Vent valve knob to OPEN.
5. Remove the vessel tray and discard the wash solvent.
Spare Parts
Part Number Description
330090 50 mL centrifuge tube, pack of 25
167550 1 L glass waste bottle (no cap)
167545 GL45 waste cap
547225 Waste Tubing Bundle
NP-1392 Cleavage vessels, pack of 120
170810 Cleavage vessel cap
170875 Vessel rack cover
Razor™ Parallel Peptide Cleavage System
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Limited Warranty
What Is Covered:
CEM Corporation warrants that the instrument will be free of any defect in parts or workmanship and will, at its option, replace or repair any defective part (excluding consumables) or instrument.
For How Long:
This warranty remains in eect for 365 days from date of delivery to the original purchaser.
What Is Not Covered:
This warranty does not cover parts or workmanship damaged due to:
Neglect, abuse or misuse, or theft
Damage caused by or to test samples,
Damage incurred during instrument relocation,
Damage caused by or to any attached equipment,
Use of incorrect line voltages or fuses,
Fire, ood, “acts of God” or other contingencies beyond the control of CEM Corporation,
Improper or unauthorized repair, or
Any other damage caused by purchaser or its agents.
Responsibilities of Purchaser:
To ensure warranty coverage, the purchaser must:
Use the instrument according to directions,
Connect the instrument properly to a power supply of proper voltage,
Replace blown fuses,
Replace consumables and
Clean the instrument as required.
How to Get Service:
Purchaser should contact the Service Department of CEM Corporation or the nearest CEM subsidiary or distributor for return authorization and for proper crating and shipping instructions to return instrument, freight prepaid, for service. On-site repairs by an authorized service technician are available through the CEM Service Department. Travel costs will be charged to the purchaser for onsite repairs.
Within the U.S. Outside the U.S.
CEM Corporation CEM Corporation 3100 Smith Farm Rd. 3100 Smith Farm Rd. Matthews, NC 28104-5044 Matthews, NC 28104-5044 (800) 726-5551 (704) 821-7015 Fax: (704) 821-4368 Fax: (704) 821-4368
Warranty Disclaimer:
CEM Corporation hereby excludes and disclaims any warranty of merchantability or tness for any particular purpose. No warranty, express or implied, extends beyond the face hereof. CEM Corporation shall not be liable for loss of use of instru­ment or other incidental or consequential costs, expenses or damages incurred by the purchaser or any other user. This warranty is not transferable.
Purchaser’s Rights under State Law:
This warranty gives the purchaser specic legal rights, and the purchaser may also have other rights that vary from state to state.
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Razor™ Parallel Peptide Cleavage System
CEM Corporation
PO Box 200
Matthews, NC 28106
Tel: 800-726-3331 Tel: 704-821-7015
Fax: 704-821-7894
Email: info@cem.com
www.cem.com
France
CEM μWave S.A.S.
Immeuble Ariane
Domaine Technologique de Saclay
4, rue Rene’ Razel
91892 ORSAY Cedex
Tel: 33 (01) 69 35 57 80 Fax: 33 (01) 60 19 64 91 Email: info.fr@cem.com
www.cemfrance.fr
Germany, Austria, Swit-
zerland
CEM GmbH
Carl-Friedrich-Gauss-Str.9
47475 Kamp-Lintfort Tel: (49) 2842-9644-0
Fax: (49) 2842-9644-11
Email: info@cem.de
www.cem.de
Ireland
CEM Technology (Ireland) Ltd.
Sky Business Centre
9a Plato Business Park
Damastown
Dublin 15
Tel: +353 (0) 1 885 1752
Fax: +353 (0) 1 885 1601
Email: info.ireland@cem.com
www.cemmicrowave.co.uk
Italy
CEM S.R.L.
Via Dell’ Artigianato, 6/8
24055 Cologno al Serio (Bg)
Tel: (39) 35-896224
Fax: (39) 35-891661
Email: info.srl@cem.com
www.cemmicroonde.com
Japan
CEM Japan K.K.
2-18-10 Takanawa
Minato-ku, Tokyo
108-0074
Tel: +81-3-5793-8542
Fax: +81-3-5793-8543
Email: info@cemjapan.jp
www.cemjapan.co.jp
CEM has been an ISO-certied
facility since 1994.
All Rights Reserved. This manual contains proprietary information which shall not be reproduced or transferred to other documents or disclosed to others without prior written permission of CEM Corporation. CEM® is a registered trademark of CEM Corporation.
www.cem.com • peptide.support@cem.com • (800) 726-3331 (US + Canada) • (704) 821-7015
Copyright 2017 by CEM Corporation.
Razor™ is a trademark of CEM Corporation.
Copyright CEM Corporation 2017 • Revision 2
United Kingdom
CEM Microwave Technology Ltd.
2 Middle Slade
Buckingham Industrial Estate
Buckingham MK181WA
Tel: (44) 1280-822873
Fax: (44) 1280-822342
Email: info.uk@cem.com
www.cemmicrowave.co.uk
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