ProZyme Gly-Q User Manual

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Gly-Q

Glycan

™

Analysis System

User Manual

Product Code: GQ2100

Version: AF

User Manual

Table of Contents

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Summary of Contents for ProZyme Gly-Q

Page 1 Gly-Q Glycan ™ Analysis System User Manual Product Code: GQ2100 Version: AF User Manual...
Page 2 ProZyme, Inc. Gly-Q™ Glycan Analysis System...
Page 3: Table Of Contents
3.9 Vacuum Manifold Setup ........................... 19 Secti on 4: Gl y-Q Mana ger Softwa re ..........................20 4.1 Starting Gly-Q Manager............................ 20 4.2 Controlling and Monitoring Gly-Q Instrument....................20 4.3 Creating a Sequence ............................21 4.3.1 Import & Export............................22 4.3.2 Sequenc e Preview .............................22 4.3.3 Autosampler Layout..........................22...
Page 4: Section 1: Safety Information
SECTION 1: SAFETY INFORMATION The Gly-Q system is intended to be installed and operated by the end user. To operate the system safely please review the following information. Warning: Both the instrument and the gel recharging air pump must be connected to an appropriate power source.
Page 5: Section 2: Introduction
SECTION 2: INTRODUCTION 2.1 The Gly-Q System The Gly-Q System is designed as a complete system for rapid analysis of oligosaccharides such as glycans. The system is comprised of capillary gel electrophoresis core integrated with a high sensitivity fluorescence detector and 96-well autosampler, replaceable capillary gel cartridge, gel recharging pump, Gly-Q Manager software and InstantQ assay kits and standards.
Page 6: Gly-Q Instrument
2.2.1 Gly-Q Instrument The Gly-Q Instrument is a compact unit comprised of a capillary gel electrophoresis core integrated with a high sensitivity fluorescence detector and 96-well autosampler. When used as part of a complete system, the instrument facilitates rapid separation, identification and relative quantitation of labeled oligosaccharides.
Page 7: Gel Recharging Air Pump
2.2.3 Cartridge Priming Station The cartridge priming station is designed to prepare the cartridge for first-time use before installing it into Gly-Q Instrument. The priming station can also be used to help troubleshoot cartridge clogging. User Manual...
Page 8: Replaceable Capillary Gel Cartridge
A pin that connects to a negatively charged electrode Label Cartridge label with lot number and expiration date Gel reservoir Top reservoir containing separation gel Guiding groove Molded groove ensures correct cartridge positioning within the instrument Cartridge tip Metal tip of the cartridge Gly-Q™ Glycan Analysis System...
Page 9: Sample Preparation And Analysis Kits
UHPLC and LC-MS applications. Gly-X with InstantQ (GX96-IPC) is specifically designed for CE separation on the Gly-Q System. Labeling (1 minute) with InstantQ dye is followed by a simple 96-well vacuum plate-based clean up step. The samples are then ready for analysis on the Gly-Q Instrument. With the deglycosylation and labeling all carried out in solution, the method is simple, rapid and suitable for automation.
Page 10: Glycan Standards
2.2.6 Glycan Standards 2.2.6.1 Gly-Q Alignment Standards Gly-Q glucose unit (GU) Ladder is generally the first run at the beginning of each sample sequence and produces >15 peaks in the electropherogram (see below). The Migration Standards (MS) co-injected with subsequent unknown samples will be aligned to the Maltodextrin (GU) ladder for glycan naming and alignment purposes.
Page 11: Gly-Q Manager Software
2.2.7 Gly-Q Manager Software The Gly-Q Manager software controls Gly-Q instrument, collects and processes data, and reports results. The software features are organized into tabs and ribbon groups for easy intuitive navigation: File Sequence Data Processing Method Instrument Method Instrument Status Bar...
Page 12 Saving an Instrument Method Instrument Instrument Status Bar informs the user: • Status Bar Instrument connectivity • Progress of run • Estimated finish time of run Note: See Section 4 for more detailed description of Gly-Q Manager functionality. Gly-Q™ Glycan Analysis System...
Page 13: Section 3: Installation
(100-240V AC, 47-63Hz). See Section 6: System Specifications. The input voltage of Gly-Q System gel recharging air pump is 12V DC and the maximum current is 6.67A. A suitable power supply with a power cord is included in the package (100-240V AC, 47-63Hz). See Section 6: System Specifications.
Page 14: System Set-Up
A surge protector is suggested to minimize power fluctuations.  Plug each power supply low voltage line into the power inlet of the appropriate unit (Gly-Q Instrument power inlet and air pump power inlet). The two connectors are different to prevent using the wrong power supply.
Page 15: Software Installation
Go through software installation steps. Note: If you have an older version of Gly-Q Manager software installed on your computer, you will be asked to uninstall it prior to installing a newer version. Installing new version of Gly -Q Manager won’t delete data.
Page 16: Cartridge Preparation
 3.7.2 Unpacking and Preparing the Cartridge  Open gel-cartridge clam-shell container and remove the cartridge  Peel off the tape from side and top of cartridge (retain top tape for storage of cartridge between uses) Gly-Q™ Glycan Analysis System...
Page 17: Priming The Cartridge
 Carefully remove and pin from package. Insert pin into the hole of the cartridge cap and push it down to the end. Slight resistance should be encountered when performing this step. 3.7.3 Priming the Cartridge  Turn off the Air Pump and use the illustration to reconnect the air tubing. ...
Page 18: Inserting The Cartridge Into The Instrument
 After gently depressing cartridge door atop Gly-Q Instrument insert cartridge and close door. Push the cartridge to the Bottom ...
Page 19: Vacuum Manifold Setup
3.9 Vacuum Manifold Setup ProZyme recommends the use of Millipore MultiScreen™ Vacuum Manifold (96 -well) and Millipore High Output Pump (WP6211560 for 115 V/60 Hz or WP6222050 for 220 V/50 Hz)     Assemble vacuum manifold.  Connect vacuum manifold and pump inlet with tubing via liquid trap reservoir.
Page 20: Section 4: Gly-Q Manager Software
SECTION 4: GLY-Q MANAGER SOFTWARE 4.1 Starting Gly-Q Manager To start the Gly-Q Manager double-click the Gly-Q Manager icon on the desktop or select Start -> Gly-Q Manager. 4.2 Controlling and Monitoring Gly-Q Instrument To control and monitor the Gly-Q Instrument with Gly-Q Manager, the instrument must be turned on and connected to a computer via USB cable.
Page 21: Creating A Sequence
4.3 Creating a Sequence A sequence defines how a group of samples is analyzed. Sequence can be created on the Sequence tab by defining the following fields: Project & Run    Name your project by selecting “New Project” from the pull down, or select from the list of existing projects.
Page 22: Import & Export
The autosampler layout will show color coded positions for samples, reagents and standards. Reagent Tray for: Park; 4 ml DI water + mineral oil to cover, optional Wash/Waste; 4 ml DI water Clean; 4 ml DI water Separation Buffer; 4 ml, included with Gly-Q Cartridge Module Gly-Q™ Glycan Analysis System...
Page 23: Running A Sequence
Sequence Status Bar An instrument status bar at the bottom of the screen will display instrument status:         Sample name  Instrument method step  Well position  Sequence progress and estimated time of completion ...
Page 24: Monitoring Sequence Progress
 The autosampler layout will show completed samples (green), failed samples (red), currently running sample (blue) and queued samples (grey).  The sequence progress table will be updated with currently running sample highlighted in darker gray. Gly-Q™ Glycan Analysis System...
Page 25: Data Review, Export, Reporting And Management
4.5 Data Rev iew, Export, Reporting and Management After data for all samples in a sequence have been acquired and processed, it is useful to review electropherograms and peak tables before reporting the results. Peak integration may need to be adjusted before results are reported.
Page 26 Note: Always start with processed data view with or without annotations. Raw data view and curren t are designed to aid in troubleshooting.  On the Sample table, select the sample(s) you want to view: Samples Gly-Q™ Glycan Analysis System...
Page 27  Open the peak table for any or all of the samples selected by clicking on sign before the sample name: Samples  Review peak detection and integration by highlighting peak line in the peak table and reviewing peak start and end markers on electropherogram view. If needed, zoom in with a left mouse click/hold; zoom out with a right mouse click and selecting un-zoom.
Page 28 To create a Virtual Data set first highlight the desired samples in the Sample table and then right click on any of the selected samples and select “Create Virtual Data Set” in the pop - up menu. Gly-Q™ Glycan Analysis System...
Page 29 A dialog opens to name the Virtual Data set. Select the desired project, enter the desired names, and if desired select a calibration ladder. Click on the “Create” button and the new Virtual Data Set will appear in the Data Files tree. Click on this data set and only the samples included in the Virtual Data Set will be in the Samples table and only the first sample will be selected and displayed.
Page 30 Export & Reporting • Selecting QDF export will create a file readable by Gly-Q Manager. This is particularly useful for sharing data with fellow researchers. • Selecting PDF export will create an electronic report that can be printed. The report will include electropherograms and peak data tables for selected samples.
Page 31: Creating A Processing Method
4.6 Creating a Processing Method Processing method defines peak alignment, peak integration and peak calling. Gly -Q Manager installs equipped with several examples of processing methods. • Enbrel_Method: contains GU values for InstantQ-labeled N-glycans from Enbrel (Fc fusion protein) produced in CHO. Mainly biantennary glycans, some with α(2,3)-linked sialic acid, with some Man5.
Page 32 Review peak detection and integration by highlighting peak line in the peak table and reviewing peak start and end markers on Electropherogram view. If needed, zoom in by clicking and dragging a rectangle around the area of interest; zoom out with a right mouse click and select un-zoom. Electropherogram Gly-Q™ Glycan Analysis System...
Page 33  In the Integration Parameters Tab there are four parameters available. • Select the Baseline Valley-to-Valley Modulation level. A setting of 100% is recommended, 0% is a flat baseline, and 50% is half way between these settings. • Select whether Detect Shoulders should be used. Checked will detect peak shoulders as separate peaks and is recommended, un-checked will identify peak with shoulders as a single peak.
Page 34: Creating An Instrument Method
 To create or modify an instrument method, select the Instrument Method tab and pick one of the existing instrument methods from the drop-down menu: I nstrument Method Tab Gly-Q™ Glycan Analysis System...
Page 35  Modify instrument commands for Calibration Ladder, Unknown Sample and End of Sequence steps by selecting Action, High Voltage, Duration and Autosampler Position: GU Ladder Samples Shutdow n The following instrument commands can be used to create an instrument method. Click on the box in the Action column in any row three times to bring up the pop-up menu below to select a command: •...
Page 36: Ending A Run
(Section 4.3.3) with water to prevent the cartridge tip from drying during storage on the instrument.  Samples may be stored, covered with sealing film, at -20°C for at least on month or up to 5 days at 4 °C. Gly-Q™ Glycan Analysis System...
Page 37: File Settings
4.9 File Settings File settings capture instrument serial number, cartridge and reagents lot information, define custom fields on data reports, perform air and gel flow checks. To update system settings, select the File tab:       ...
Page 38  Account settings:  Data storage settings: Note: This setting is for information only and cannot be changed.  Airflow check:  Gel Flow check: Gly-Q™ Glycan Analysis System...
Page 39: Section 5: Care And Maintenance
Check the air filter. If the filter is damaged or wet, use a new filter to replace the old one. Replacement air filters are supplied with the instrument and are available from ProZyme. • Turn on the power to the air pump; it will run briefly to pressurize the system and then turn off.
Page 40: Section 6: System Specifications
MEAN WELL GSM90B12-P1M Power Supply Input Voltage 100 – 240 V AC, 47-63 Hz Suggestion: Use a surge protector outlet for the instrument and the computer to avoid electromagnetic interferences and damages to the inner electrical devices. Gly-Q™ Glycan Analysis System...
Page 41: Section 7: Warranty Statement
SECTION 7: WARRANTY STATEMENT Limitation of Liability Gly-Q Analyzer, software, and all related reagents are designed for the use of electrophoresis analysis in general bio-chemistry laboratory for Research Use Only. BEFORE ATTEMPTING TO OPERATE THE INSTRUMENT, READ ALL PRODUCT MANUALS AND FOLLOW THE INSTRUCTIONS.
Page 42: Section 8: Instructions For Depot Repair Returns
 Print and fill out the Decontamination Certificate located on the next page of this User Manual. Pack the form with the instrument and contact ProZyme or your local distributor for shipping details. The User Manual is available at http://prozyme.com/products/gq2100...
Page 43 Decontamination Certificate Please complete the form carefully and attach it to the item or instrument being returned Return authorization number Company name Contact person Contact phone number Instrument model Instrument serial number List accessories returned with the instrument  No decontamination required This instrument, and accessories if applicable, has not been in contact with any biological, chemical, or radioactive material that might cause infection or injury.
Page 44: Section 9: Faqs
Does the Gly-Q instrument require a minimum volume for analysis? The Gly-Q instrument requires a volume of at least 30 µL in a 96 well plate well for a run. Gly-Q Manager does not seem to be communicating to the instrument. How can this be fixed? Ensure that the USB cable is intact and installed correctly.
Page 45 This message indicates that there is a problem reading the information on the cartridge RFID tag. In most cases the system will allow you to enter the cartridge information manually (see section 4.4 .1) but in this case manual entry is no longer working. If this happens you must contact ProZyme technical support for assistance.
Page 46 Sample. Please see below for an example. Manually designated LMS or UMS will be named accordingly in bold print, see below: Gly-Q™ Glycan Analysis System...
Page 47 You can move data to a different computer running Gly-Q Manager using the data export and data import functions using the QDF data file format. If you want to analyze data using programs that support the industry standard AIA format such as Chromeleon or Empower you can use the data export function with the CDF data file format.
Page 48: Section 10: Legal Notices
Inc., in the United States and other countries. All other trademarks are the property of their respective owners. Failure of a mark to appear on this page does not mean that ProZyme does not use the mark nor does it mean that the product is not actively marketed nor is not significant within its relevant market.
Page 49: Section 11: Resources And References
SECTION 11: RESOURCES AND REFERENCES Visit ProZyme’s website for additional information, downloadable posters, publications and technotes. http://prozyme.com/pages/resources User Manual...
Page 50: Section 12: Technical Assistance
SECTION 12: TECHNICAL ASSISTANCE ProZyme is committed to developing rapid, automatable methods for glycan analysis. Call us to discuss products in development. If you have any questions or experience difficulties regarding any aspect of our products, please contact us: TOLL FREE (800) 457-9444 (US & CANADA)
Page 51 3832 Bay Center Place N-Glycanase, Gly-Q and InstantQ are trademarks or registered Hayward, CA 94545 trademarks of ProZyme, Inc., in the United States and other countries. Toll Free: +1 (800) 457-9444 All other trademarks are the property of their respective owners.