41-0162-PD - BLItz IQOQ - Manual - Guide

BLITZ® SYSTEM IQOQ USER MANUAL
INSTALLATION & OPERATION QUALIFICATION PROTOCOLS

© 2013 ForteBio, Inc. All rights reserved.
No part of this publication may be reproduced, transmitted, transcribed, stored in retrieval systems, or translated into any
form, or by any means: electronic, mechanical, magnetic, optical, or otherwise, without the prior written permission of
ForteBio, 1360 Willow Road, Suite 205, Menlo Park, CA 94025, United States of America.
Disclaimer
ForteBio reserves the right to change its products and services at any time to incorporate the latest technological
developments. This user manual is subject to change without notice.
Limitation
The ForteBio BLItz system is for research use only, not for use in diagnostic procedures.
Trademarks
BLItz is a registered trademark of ForteBio, Inc.
P/N 41-0162-PD Rev A
III
BLITZ SYSTEM IQOQ
TABLE OF CONTENTS
Table of Contents ...........................................................................................................................................................IV
SECTION 1: Qualification Description for the BLItz System .......................................................................................... 2
1.1. Purpose ...............................................................................................................................................................2
1.2. Scope...................................................................................................................................................................2
1.3. Introduction ........................................................................................................................................................2
1.4. BLItz System Overview ........................................................................................................................................2
1.5. Comments (page intentionally left blank) ..........................................................................................................4
1.6. Pre-Approvals......................................................................................................................................................5
SECTION 2: Installation Qualification of the BLItz System ............................................................................................ 6
2.1. Purpose ...............................................................................................................................................................6
2.2. Materials Required to Install the BLItz System ...................................................................................................6
2.3. System Owner .....................................................................................................................................................6
2.4. Receiving Inspection ...........................................................................................................................................6
2.5. Comments (page intentionally left blank) ..........................................................................................................9
2.6. Setting up the BLItz System ..............................................................................................................................10
2.7. Communication Verification .............................................................................................................................12
2.8. Software Verification ........................................................................................................................................12
2.9. Documentation .................................................................................................................................................12
2.10. Comments (page intentionally left blank) ........................................................................................................13
2.11. Installation Qualification Sign Off ..................................................................................................................... 14
SECTION 3: Operation Qualification of the BLItz System ............................................................................................ 15
3.1. Purpose .............................................................................................................................................................15
3.2. Method .............................................................................................................................................................15
3.3. Materials Required For Operation Qualification ..............................................................................................15
3.4. Quantitation Qualification ................................................................................................................................16
IV
3.4.1. Overview of the Method ..........................................................................................................................16
3.4.2. Sample Preparation .................................................................................................................................17
3.4.3. Data Acquisition (Experiment Set Up and Run) Protocol .........................................................................17
3.4.4. Data Analysis Protocol .............................................................................................................................21
3.4.5. Repeat Quantitation Qualification ...........................................................................................................22
3.5. Kinetics Qualification ........................................................................................................................................23
3.5.1. Overview of the Method ..........................................................................................................................23
3.5.2. Sample Preparation .................................................................................................................................23
3.5.3. Data Acquisition (Experiment Set Up and Run) Protocol .........................................................................24
3.5.4. Data Analysis Protocol .............................................................................................................................29
3.5.5. Repeat Kinetics Qualification ...................................................................................................................29
3.6. Comments (page intentionally left blank) ........................................................................................................30
3.6. Operation Qualification Sign Off ....................................................................................................................... 31
V
BLITZ SYSTEM IQOQ
SECTION 1: QUALIFICATION DESCRIPTION FOR THE BLITZ SYSTEM
1.1. PURPOSE
The purpose of the BLItz system Installation Qualification and Operation Qualification (IQOQ) user manual is to
provide a guide to verify proper installation and operation of the BLItz system.
1.2. SCOPE
This document provides detailed steps to perform an IQOQ of the BLItz system. System performance specific to an
end-user application is considered to be a Performance Qualification (PQ) and outside of the scope of the IQOQ.
Validation of the BLItz Pro software and Dip and Read™ biosensors are also outside the scope of the IQOQ process.
The BLItz system IQOQ Validation and Testing kit is intended for use in qualifying a
NOTE single BLItz system. An IQOQ package should be purchased for each additional BLItz
system to be qualified.
1.3. INTRODUCTION
The BLItz system IQOQ user manual includes three sections:
SECTION DESCRIPTION
SECTION 1: QUALIFICATION DESCRIPTION DESCRIPTION OF THE OVERALL VALIDATION PROCEDURE

FOR THE BLITZ SYSTEM FOR THE BLITZ SYSTEM
SECTION 2: INSTALLATION DOCUMENTATION AND VERIFICATION OF PROPER

QUALIFICATION OF THE BLITZ SYSTEM INSTALLATION OF THE BLITZ SYSTEM
SECTION 3: OPERATION QUALIFICATION DOCUMENTED QUALIFICATION OF BLITZ SYSTEM
OF THE BLITZ SYSTEM OPERATION TO FACTORY SPECIFICATIONS
1.4. BLITZ SYSTEM OVERVIEW
The BLItz system comes with a power supply, connection cables, and a starter kit. BLItz Pro™ software for data
acquisition and analysis is provided on a CD.
The BLItz system utilizes ForteBio’s proprietary Bio-layer Interferometry (BLI) technology to monitor biomolecular
interactions in real time without the use of labels. Using this optical technique, binding between ligands
immobilized on the tip of a biosensor and an analyte in solution is measured. A change in optical thickness at the
2
biosensor tip results in a proportional wavelength shift at the detector. Sample analysis is performed in either the
BLItz sample drop holder or a microfuge tube.
The BLItz system user interface is based on five Experiment types:
• Quick Yes/No: tests samples for the presence of a protein.
• Create Standard Curve and Quantitate Sample: provide a measure of active protein concentration in
solution.
• Basic Kinetics and Advanced Kinetics: measure the association (ka) and dissociation (kd) constants, and
affinity (KD) for binding interactions.
In a Quick Yes/No Experiment, a negative control and positive control sample are tested along with an unknown
sample. The presence of protein is determined if the unknown displays a binding signal.
In a Create Standard Curve Experiment, each distinct sample concentration produces a unique binding rate. A set
of standards with known concentrations is used to generate a standard curve of binding rates vs. concentrations.
The sample binding rate is compared to the standard curve to derive accurate quantitation information in the
Quantitate Sample module. After the quantitation data is analyzed, a report can be generated that displays the
setup, standard curve, and calculated concentrations.
The Basic Kinetics module is appropriate for use when the ligand has previously been loaded on the biosensor.
Baseline and dissociation steps are performed in a microcentrifuge tube, and the association step is performed in
either the drop holder or the tube. If the duration of the association step is more than five minutes, the drop
holder should not be used.
The Advanced Kinetics module can be used to perform kinetic analysis of ligand-analyte interactions where the
ligand is first loaded on the biosensor in the BLItz system, followed by monitoring the association and dissociation
of analyte to the ligand. The Advanced Kinetics module can also be used to set up custom binding assays involving
multiple binding steps, such as the formation of a sandwich complex (capture antibody-antigen-secondary
antibody).
Binding curves are generated in real time during a run and displayed in the Runtime Binding Chart window in BLItz
Pro software.
For more detailed information on running Experiments, please refer to the User Guide provided in the Help menu
of BLItz Pro software.
3
BLITZ SYSTEM IQOQ
1.5. COMMENTS (PAGE INTENTIONALLY LEFT BLANK)
4
1.6. PRE-APPROVALS
By signing this page, the following individuals indicate that they have reviewed and agreed to the IQOQ procedure
prior to the commencement of qualification.
AUTHORIZED CUSTOMER REPRESENTATIVE
NAME:
TITLE:
SIGNATURE: DATE:
NAME:
TITLE:
SIGNATURE: DATE:
NAME:
TITLE:
SIGNATURE: DATE:
5
BLITZ SYSTEM IQOQ
SECTION 2: INSTALLATION QUALIFICATION OF THE BLITZ SYSTEM
2.1. PURPOSE
The BLItz system Installation Qualification is intended to verify that the correct system was ordered, delivered, and
installed according to ForteBio specifications. If the qualification is being performed during the installation of the
BLItz system, please use the procedure described in this section to perform the installation and to record the
process. For BLItz systems previously installed, please use this section to record and verify details of the
installation.
2.2. MATERIALS REQUIRED TO INSTALL THE BLITZ SYSTEM
The user should provide a laptop or desktop computer to operate the BLItz system. The minimum computer
specifications are:
• 2.5 GHz Dual Core CPU
• 2 GB RAM
• 32-bit or 64-bit Windows XP and Windows 7
• One (1) USB 2.0 port
• One (1) monitor (minimum 1,024 X 768)
2.3. SYSTEM OWNER
Record the name, institution and location information for the primary user of the BLItz system.
NAME:
INSTITUTION:
LOCATION:
2.4. RECEIVING INSPECTION
1. Unpack the box.
2. Inspect all the contents. Using the checklists in Tables 2.1 and 2.2, confirm that all of the required components
are included.
6
TABLE 2.1 BLITZ SYSTEM PACKAGE CHECKLIST
ITEM CHECK IF RECEIVED
BLITZ INSTRUMENT
BLITZ PRO SOFTWARE CD
POWER CORD
USB CABLE
BLITZ STARTER KIT
TABLE 2.2 BLITZ STARTER KIT CHECKLIST
ITEM CHECK IF RECEIVED
ONE TRAY OF 32 PROTEIN A BIOSENSORS
200 µL OF 10 mg/mL HUMAN IGG
100 µL OF 10 mg/mL MOUSE IGG
50 mL OF SAMPLE DILUENT
500 µL OF 0.5 M HCL
ONE DROP HOLDER
TWO 96-WELL PLATES
20 MICROCENTRIFUGE TUBES (0.5 mL)
SWAB, PACK OF 20
SILVER SHARPIE MARKER
BLITZ QUICKSTART GUIDE
7
BLITZ SYSTEM IQOQ
3. Record the serial number of the BLItz instrument. This can be found on the product label located at the rear
of the instrument.
SERIAL NUMBER
4. Record the lot numbers of the consumables from the Starter Kit. These can be found on the product labels.
ITEM LOT NUMBER
ONE TRAY OF 32 PROTEIN A BIOSENSORS
200 µL OF 10 mg/mL HUMAN IGG
100 µL OF 10 mg/mL MOUSE IGG
50 mL OF SAMPLE DILUENT
500 µL OF 0.5 M HCL
5. Inspect the following to verify:
ITEM CHECK IF VERIFIED
THE BLITZ SERIAL NUMBER MATCHES THAT LISTED

ON THE INVOICE.
THE BLITZ IS FREE OF PHYSICAL DAMAGE.
THE SLIDER MOVES FREELY FROM LEFT TO RIGHT

(SEE FIGURE 2).
THE COVER OF THE BLITZ INSTRUMENT CLOSES
PROPERLY, IN BOTH TUBE AND DROP HOLDER
READING POSITIONS (FIGURE 2).
THE ARM DROPS AFTER CLOSING THE LID OF THE
BLITZ SYSTEM.
6. Document any missing items or damage. If you take any corrective actions, record them in the following
Comments section.
8
9
BLITZ SYSTEM IQOQ
2.6. SETTING UP THE BLITZ SYSTEM
1. Place the BLItz system on a dry, level surface with minimal vibration and away from direct sunlight.
2. Insert the BLItz Pro software CD in a laptop or desktop computer and run the BlitzInstallCD.exe file.
3. Click Install BLItz Pro Software (Figure 1) to install the software.
4. Click Install BLItz Driver (Figure 1) to install the required hardware driver support.
FIGURE 1. BLITZ PRO SOFTWARE INSTALLTION SCREEN.
7. Click Exit to complete the software installation.
8. Connect the power cord to the BLItz system and plug the other side into an outlet (Figure 3).
10
9. Connect one side of the data cord to the BLItz system and the other to a USB port on your computer (Figure
3). Not all USB ports, especially on laptop computers, are fully powered, so connecting to a USB port at the
rear on the computer is recommended.
FIGURES 2 AND 3. FRONT AND REAR VIEWS OF THE BLITZ SYSTEM.
10. Place a drop holder in its slot on the BLItz system (Figure 4).
FIGURE 4. PLACING THE DROP HOLDER INTO THE BLITZ SYSTEM.
11
BLITZ SYSTEM IQOQ
2.7. COMMUNICATION VERIFICATION
1. Power on the BLItz system and wait for the Windows® plug-and-play manager to assign drivers.
2. Confirm that the BLItz system’s front light comes on.
3. Double-click the BLItz Pro desktop icon.
4. Verify that the following actions have taken place:
ITEM CHECK IF VERIFIED
AN “INITIALIZING” PROMPT DISPLAY
THE BLITZ SYSTEM’S FRONT LIGHT BLINKS
THE “INITIALIZING” PROMPT CLOSES

AUTOMATICALLY AND THE LIGHT HAS STOPPED
BLINKING
2.8. SOFTWARE VERIFICATION
1. In BLItz Pro software, select Help > About BLItz... on the menu bar.
2. In the pop-up screen that appears, find and record the software version number. Ensure that this version
number matches that shown in the title bar.
BLITZ PRO SOFTWARE VERSION NUMBER
2.9. DOCUMENTATION
Hard copies of the BLItz User Guides are not included but can be found as electronic PDFs in the Help menu of
BLItz Pro software.
ITEM CHECK IF AVAILABLE
BLITZ SYSTEM USER GUIDE
12
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BLITZ SYSTEM IQOQ
2.11. INSTALLATION QUALIFICATION SIGN OFF
All steps required for the BLItz system Installation Qualification have been completed, documented and reviewed.
All discrepancies noted during Installation Qualification have been satisfactorily resolved. The installation of the
BLItz system is hereby considered qualified.
NAME:
TITLE:
SIGNATURE: DATE:
NAME:
TITLE:
SIGNATURE: DATE:
NAME:
TITLE:
SIGNATURE: DATE:
14
SECTION 3: OPERATION QUALIFICATION OF THE BLITZ SYSTEM
3.1. PURPOSE
The purpose of the operation qualification protocol is to verify that:
• The BLItz system is fully functional after the installation.
• System functionality is as expected for both quantitation and kinetics analyses.
• Consistent and reproducible results are obtained and are within product specifications.
3.2. METHOD
The Operation Qualification (OQ) of the BLItz system is performed in two stages:
• Quantitation Qualification (QQ)
• Kinetics Qualification (KQ)
ForteBio recommends performing the quantitation qualification first followed by the

NOTE
kinetics qualification.
3.3. MATERIALS REQUIRED FOR OPERATION QUALIFICATION
All reagents, biosensors, and sample vessels required for the qualification are provided in the BLItz Validation and
Testing kit.
• IQOQ CD
• BLItz system IQOQ User Manual
• Protein A biosensors – 1 tray of 32 biosensors
• Human IgG – 1 tube of 10 mg/mL
• Mouse IgG – 1 tube of 10 mg/mL
• Hydrochloric acid – 1 tube of 0.5 M
• Sample Diluent – 1 bottle
• Drop holder
15
BLITZ SYSTEM IQOQ
ForteBio recommends cleaning the drop holders prior to using the BLItz system for the
first time. Rinse each drop holder once with 10 µL of 0.5 M hydrochloric acid followed
NOTE
by three rinses with at least 10 µL of Sample Diluent. After each rinse, wipe the drop
holder with a swab or Kimwipe®.
• 96-well, black, flat-bottom, polypropylene (Greiner Bio-one, #665209) microplates – 2 microplates
• 0.5 mL, black, microcentrifuge tubes – 20 tubes
• Swab – Pack of 20
The following materials are required for the Operation Qualification, but are not provided in the BLItz Validation
and Testing kit. These materials should be provided by the customer:
• Micropipettors, various standard volumes
• Pipette tips, various standard sizes
• Sharps waste container
3.4. QUANTITATION QUALIFICATION
Quantitation qualification is performed using Protein A biosensors to measure high and low concentration samples
of human IgG.
For best results, the BLItz system should be powered on with the cover closed for at
NOTE
least one hour before starting this test.
3.4.1. Overview of the Method
• Prepare two samples of human IgG (100 and 1000 µg/mL concentrations).
• Hydrate six Protein A biosensors for use.
• Quantitate six hIgG samples.
• Calculate the initial binding rates of the hIgG samples.
A criterion of %CV ≤ 10% for 100 µg/mL and ≤ 12% for 1000 µg/mL is employed to evaluate PASS or FAIL of the
BLItz system for quantitation qualification.
16
The Quantitation Qualification should be performed only with reagents and

NOTE
biosensors provided with the BLItz Validation and Testing kit.
3.4.2. Sample Preparation
1. Allow all reagents to warm to room temperature.
2. Prepare the 1000 µg/mL human IgG sample by pipetting 10 µL of the stock 10 mg/mL hIgG into 90 µL of
sample diluent. Mix by gently pipetting up and down.
3. Prepare the 100 µg/mL hIgG sample by pipetting 10 µl of the 1000 µg/mL solution prepared in the previous
step into 90 µL of sample diluent. Mix by gently pipetting up and down.
NOTE All dilutions should be made using the provided Sample Diluent only.
3.4.3. Data Acquisition (Experiment Set Up and Run) Protocol
1. Set up the experiment in BLItz Pro software:
Use the method file provided on the CD for proper execution of the qualification
NOTE protocol. If you are unable to access the files from the BLItz IQOQ CD, contact
ForteBio Technical Support at support@fortebio.com.
1.1. Ensure that the instrument has been on for at least one hour.
1.2. Load the IQOQ CD into the computer controlling the BLItz system.
1.3. Click on File in the menu options in BLItz Pro software and choose Open....
1.4. Locate the IQOQ_Quant.dataset method file:
1.4.1. On the IQOQ CD, go to the METHOD FILES folder.
1.4.2. Click on IQOQ_Quant.dataset.
2. Hydrate six biosensors:
Biosensors should be hydrated for at least 10 minutes before use in any

IMPORTANT
experiment.
17
BLITZ SYSTEM IQOQ
• Hydrate only those sensors to be used in an experiment. 6 are needed for the Quantitation
Qualification.
• Hydrate just before running an experiment.
• Hydrate for at least 10 minutes.
• Do not allow the hydrated biosensors to dry. Load the biosensor onto the BLItz system only when
prompted to do so by the software.
2.1. Add 200 μL of sample diluent to the wells of a 96-well microplate that corresponds to the biosensors that
will be used in the experiment. For example, if using biosensors A1 to A6 from the biosensor tray, add
buffer only to wells A1 to A6 in the 96-well microplate.
2.2. Remove the clear cover of the Protein A biosensor tray.
2.3. Carefully lift the green biosensor rack out of the blue biosensor tray holder (Figure 5) - avoiding touching
of the biosensor tips on any solid surface.
FIGURE 5. REMOVE THE GREEN BIOSENSOR RACK FROM THE BLUE TRAY HOLDER TO INSERT
HYDRATION MICROPLATE.
2.4. Place the hydration microplate inside the blue biosensor tray holder.
2.5. Carefully lower the green biosensor rack on top of the hydration microplate in the biosensor tray holder
to begin hydrating the biosensors.
2.6. Select a hydration time of 10 minutes (the default) or more in BLItz Pro software, and click the Hydrate
Biosensor icon (Figure 6).
FIGURE 6. SETTING THE HYDRATION TIME IN THE SOFTWARE.
18
Other lab stopwatch methods can be used to monitor the hydration time outside of
NOTE
the software.
2.7. After the timer reaches zero, the icon changes to Hydration Done. The window shakes until the
Hydration Done icon is clicked (Figure 7).
FIGURE 7. HYDRATION DONE ICON APPEARS AFTER THE COUNTDOWN IS COMPLETED.
After hydrating, leaving a biosensor on the biosensor mount or exposed to air to dry
NOTE will adversely affect its performance. If you do not plan to perform an experiment
immediately, leave the biosensors in the hydration solution.
3. Start the experiment.
Ensure the biosensors have been hydrated for at least 10 minutes prior to starting
IMPORTANT
the experiment.
3.1. Verify the Run Settings are correct.
• Sample ID: Run 1 - Human IgG (100 µg/mL)
• Biosensor type: Protein A
• Concentration units: µg/mL
• Run time (s): 60
• Shaker: Enable
3.2. Click Next, and follow the on-screen instructions.
3.3. Add 4 μL of 100 µg/mL hIgG into the drop holder for the first run(Figure 8).
19
BLITZ SYSTEM IQOQ
FIGURE 8. PIPET THE SAMPLE DIRECTLY INTO THE DROP HOLDER.
3.4. Load the Protein A biosensor into the biosensor mount and give it a final quarter-turn upward twist to
ensure a snug fit (Figure 9). When handling the biosensor, do not allow the biosensor surface to come in
contact with anything.
FIGURE 9. ATTACH THE BIOSENSOR TO THE MOUNT BY PUSHING UPWARD.
3.5. Move the slider to its right-most position to move the drop holder to the read position if it is not there
already.
3.6. Close the BLItz system cover to start data acquisition. Data is collected in real time and shown on-screen
as soon as the cover is closed.
3.7. After the run, save the experiment file by navigating to File > Save As. Enter a file name, such as “BLItz
Quant OQ” and save the dataset.
3.8. Lift the cover, discard the used biosensor in a sharps container, wipe the drop holder with a swab or
Kimwipe, and rinse three times with at least 10 µL of sample diluent, wiping after each rinse with a swab
or Kimwipe.
3.9. Perform subsequent sample runs by repeating the steps above for each sample. In total, there will be
three replicates each of 100 µg/mL and 1000 µg/mL hIgG concentrations. It is recommended that the ID
for each Sample be changed in the Run Settings for each sample that is run. Examples of a naming
scheme may be:
• Run 1 - Human IgG (100 µg/mL)
20
3.10. Record the binding rates in the table below.
RUN BINDING RATE (NM/S)
RUN 1 - HUMAN IGG (100 µg/mL)
3.4.4. Data Analysis Protocol
1. Locate the IQOQ_Calculations.xlsx file:
1.1. On the IQOQ CD, go to the RESULTS folder.
1.2. Click on IQOQ_Calculations.xlsx.
2. In the spreadsheet, select the Paste Quant Raw Data here tab.
3. In BLItz Pro software, go to the Run List section of the Quantitation Qualification (Figure 10).
4. Select and highlight the values in the Binding rate (nm/s) column (Figure 10)
FIGURE 10. SELECT THE BINDING RATE DATA IN THE RUN LIST SECTION.
5. Right-click on the highlighted values and choose Copy Selected.
21
BLITZ SYSTEM IQOQ
6. In the IQOQ_Calculations.xlsx spreadsheet, click in cell B2 of the Paste Quant Raw Data here tab and paste the
copied binding rates into the cell.
7. Results are automatically calculated on the Results Summary tab.
3.4.5. Repeat Quantitation Qualification
If the BLItz system passed quantitation qualification, a repeat test is not needed and you can proceed to the
Kinetics Qualification.
In the event of a failed quantitation qualification, consult with a ForteBio Technical Support representative to
determine if a repeat test is required. The BLItz IQOQ Validation and Testing kit contains enough biosensors,
reagents and microplates to perform a repeat test. Do not proceed to the Kinetics Qualification unless instructed
by a ForteBio technical representative.
22
3.5. KINETICS QUALIFICATION
Kinetics qualification is performed using Protein A biosensors to assay mouse IgG samples.
For best results, the BLItz system should be powered on with the cover closed for at
NOTE
least one hour before starting this test.
ForteBio recommends thoroughly cleaning the drop holder before starting the kinetics
IMPORTANT qualification. Rinse once with 10 µL of 0.5 M hydrochloric acid followed by three rinses
with at least 10 µL of Sample Diluent. After each rinse, wipe the drop holder with a
swab or Kimwipe.
3.5.1. Overview of the Method
• Prepare one sample of mouse IgG (50 µg/mL concentration).
• Hydrate four Protein A biosensors for use.
• Measure the binding rate of duplicate blank samples and duplicate mouse IgG samples.
• Generate the equilibrium dissociation constant (KD) of the mIgG sample.
A PASS result is obtained when the KD is between 2.3E-9 - 1.1E-8 M.
The Kinetics Qualification should be performed only with reagents, biosensors and
NOTE
black tubes provided with the BLItz Validation and Testing kit.
3.5.2. Sample Preparation
1. Allow all reagents to warm to room temperature.
2. Prepare the 1000 µg/mL mouse IgG sample by pipetting 10 µL of the stock 10 mg/mL mIgG into 90 µL of
sample diluent. Mix by gently pipetting up and down.
3. Prepare the 50 µg/mL mIgG sample by pipetting 5 µL of the 1000 µg/mL solution prepared in the previous step
into 95 µL of sample diluent. Mix by gently pipetting up and down.
NOTE All dilutions should be made using the provided Sample Diluent only.
23
BLITZ SYSTEM IQOQ
3.5.3. Data Acquisition (Experiment Set Up and Run) Protocol
1. Set up experiment in BLItz Pro software
Use the method file provided on the CD for proper execution of the qualification
NOTE protocol. If you are unable to access the files from the BLItz IQOQ CD, contact
ForteBio Technical Support at support@fortebio.com.
1.1. Ensure that the instrument has been on for at least one hour.
1.2. Load the IQOQ CD into the computer controlling the BLItz system.
1.3. Click on File in the menu options in BLItz Pro software and choose Open....
1.4. Locate the IQOQ_Kinetics.dataset method file:
1.4.1. On the IQOQ CD, go to the METHOD FILES folder.
1.4.2. Click on IQOQ_Kinetics.dataset.

2. Hydrate four biosensors:
Biosensors should be hydrated for at least 10 minutes before use in any

IMPORTANT
experiment.
• Hydrate only those sensors to be used in an experiment. 4 are needed for the Quantitation
Qualification.
• Hydrate just before running an experiment.
• Hydrate for at least 10 minutes.
• Do not allow the hydrated biosensors to dry. Load the biosensor onto the BLItz system only when
prompted to do so by the software.
2.1. Add 200 μL of sample diluent to the wells of a 96-well microplate that correspond to the biosensors that
will be used in the experiment. For example, if using biosensors A1 to A4 from the biosensor tray, add
buffer only to wells A1 to A4 in the 96-well microplate.
2.2. Remove the clear cover of the Protein A biosensor tray.
2.3. Carefully lift the green biosensor rack out of the blue biosensor tray holder (Figure 11) - avoiding
touching of the biosensor tips on any solid surface.
24
FIGURE 11. REMOVE THE GREEN BIOSENSOR RACK FROM THE BLUE TRAY HOLDER TO INSERT
HYDRATION MICROPLATE.
2.4. Place the hydration microplate inside the blue biosensor tray holder.
2.5. Carefully lower the green biosensor rack on top of the hydration microplate in the biosensor tray holder
to begin hydrating the biosensors.
2.6. Select a hydration time of 10 minutes (the default) or more in BLItz Pro software, and click the Hydrate
Biosensor icon (Figure 12).
FIGURE 12. SETTING THE HYDRATION TIME IN THE SOFTWARE.
Other lab stopwatch methods can be used to monitor the hydration time outside of
NOTE
the software.
2.7. After the timer reaches zero, the icon changes to Hydration Done. The window shakes until the
Hydration Done icon is clicked (Figure 13).
FIGURE 13. HYDRATION DONE ICON APPEARS AFTER THE COUNTDOWN IS COMPLETED.
25
BLITZ SYSTEM IQOQ
After hydrating, leaving a biosensor on the biosensor mount or exposed to air to dry
NOTE will adversely affect its performance. If you do not plan to perform an experiment
immediately, leave the biosensors in the hydration solution.
3. Start the experiment.
Ensure the biosensors have been hydrated for at least 10 minutes prior to starting
IMPORTANT
the experiment.
3.1. Verify the Run Settings are correct.
• Sample ID: Run 1 - Blank
• Biosensor type: Protein A
• Shaker: Enable
• Concentration: 0 µg/mL
• Molecular Weight: 150 kDa
• Molar Concentration: 0 nM
• Baseline: 30 s: Tube
• Association: 120 s: Drop
• Disassociation: 180 s: Tube
3.2. Click Next, and follow the on-screen instructions.
3.3. Add 250 μL of the sample diluent to a microcentrifuge tube and insert it into the sample block.
3.4. Load the Protein A biosensor into the biosensor mount and give it a final quarter-turn upward twist to
ensure a snug fit (Figure 14).
FIGURE 14. ATTACH THE BIOSENSOR TO THE MOUNT BY PUSHING UPWARD.
3.5. Move the slider to its left-most position so that the biosensor will insert into the microcentrifuge tube
when the cover is closed.
26
3.6. Close the BLItz system cover to start data acquisition.
3.7. After the baseline is acquired, you will be prompted to open the cover to pipette 4 µL of sample diluent
into the drop holder. Move the sample stage to the right-most position to read from the drop holder
position, and close the cover.
Do not allow the biosensor surface to be exposed to air for extended periods of time
NOTE
between the steps in the kinetics assay.
3.8. Once the association step is complete, you will be prompted again to open the cover, move the slider
back to the left-most tube reading position, and close the cover.
3.9. After the run is complete, save the experiment file by navigating to File > Save As. Enter a file name,
such as “BLItz Kinetic OQ” and save the dataset.
3.10. Lift the cover, discard the used biosensor in a sharps container and microcentrifuge tube, wipe the drop
holder with a swab or Kimwipe, and rinse 3 times with at least 10 µL of sample diluent, wiping after each
rinse with a swab or Kimwipe.
3.11. Perform subsequent sample runs by repeating the steps above to obtain duplicate measurements for the
Blank sample and the 50 µg/mL hIgG sample. It is recommended that the ID for each Sample be changed
in the Run Settings for each sample that is run. Be sure to update the concentration in the Run Settings
for the Mouse IgG samples. Examples of a naming scheme may be:
• Run 1 - Blank
• Run 2 - Blank
• Run 3 - Mouse IgG (50 µg/mL)
• Run 4 - Mouse IgG (50 µg/mL)
3.12. Select a blank from Run 1 or Run 2 to use as a reference in subsequent analysis. A proper reference
should be a relatively flat horizontal line as seen in the Run Data window (Figure 15). To select a
reference, click in the checkbox of the Ref column (Figure 16).
FIGURE 15. INSPECT RUNS 1 AND 2 TO CHOOSE A SUITABLE REFERENCE. A PROPER

REFERENCE SHOULD BE A RELATIVELY FLAT HORIZONTAL LINE.
27
BLITZ SYSTEM IQOQ
FIGURE 16. TO SELECT A REFERENCE, CLICK THE CHCKBOX IN THE REF COLUMN.
3.13. Click Analyze in the Analysis Data window (Figure 17) and record the KD for the 50 µg/mL samples in the
following table. With a Global Fitting model, the reported KD values should be identical.
FIGURE 17. CLICK ON THE ANALYZE BUTTON IN THE ANALYSIS DATA WINDOW TO GENERATE KD
VALUES. ENSURE THAT THE APPROPRIATE STEP CORRECTION AND FITTING OPTIONS HAVE BEEN
SELECTED.
IS VALUE BETWEEN
RUN K D (M)
2.3E-9 - 1.1E-8 M
RUN 3 - MOUSE IGG (50 µg/mL)
RUN 4 - MOUSE IGG (50 µg/mL)
28
3.5.4. Data Analysis Protocol
There are two qualification criteria for kinetics: the calculated KD value and the RMS noise.
1. If the KD values recorded above are between 2.3E-9 and 1.1E-8 M, the calculated KD value passes that
qualification criteria.
2. To calculate the RMS noise, locate the IQOQ_Calculations.xlsx file:
2.1. On the IQOQ CD, go to the RESULTS folder.
2.2. Click on IQOQ_Calculations.xlsx.
3. In the spreadsheet, select the Paste Kinetics Raw Data here tab.
4. In BLItz Pro software, select File > Export to CSV.
5. You will be prompted to save to a folder. Create a new folder to avoid deleting existing data.
6. Navigate to the contents of the folder using Windows Explorer. There will be four .csv files in the folder. The
files will have the same name as your Kinetics Qualification data file, with _001, 002, 003, and 004 appended
to the file name. The 001 and 002 files should correspond to your sample blanks.
7. Open the _001 appended file, then highlight and copy columns A and B.
8. In the IQOQ_Calculations.xls spreadsheet, paste the copied columns into columns B and C of the Paste Kinetics
Raw Data here tab.
9. Open the _002 appended file, then highlight and copy columns A and B.
10. In the IQOQ_Calculations.xls spreadsheet, paste the copied columns into columns D and E of the Paste Kinetics
Raw Data here tab.
11. Results are calculated automatically on the Results Summary tab.
3.5.5. Repeat Kinetics Qualification
If the BLItz system passed kinetics qualification, a repeat test is not needed. You have completed the operational
qualification of the system. In the event of a failed kinetics qualification, consult with a ForteBio Technical Support
representative to determine if a repeat test is required. The BLItz IQOQ Validation and Testing kit contains enough
biosensors, reagents and microplates to perform a repeat test.
29
BLITZ SYSTEM IQOQ
30
3.6. OPERATION QUALIFICATION SIGN OFF
All steps required for the BLItz system Operation Qualification have been completed, documented and reviewed.
All discrepancies noted during Operation Qualification have been satisfactorily resolved. The BLItz system is
qualified for use.
NAME:
TITLE:
SIGNATURE: DATE:
NAME:
TITLE:
SIGNATURE: DATE:
NAME:
TITLE:
SIGNATURE: DATE:
31

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BLITZ® SYSTEM IQOQ USER MANUAL

INSTALLATION & OPERATION QUALIFICATION PROTOCOLS

BLItz is a registered trademark of ForteBio, Inc.

P/N 41-0162-PD Rev A

Table of Contents ...........................................................................................................................................................IV

SECTION 1: Qualification Description for the BLItz System .......................................................................................... 2

1.1. Purpose ...............................................................................................................................................................2

1.3. Introduction ........................................................................................................................................................2

1.4. BLItz System Overview ........................................................................................................................................2

1.5. Comments (page intentionally left blank) ..........................................................................................................4

SECTION 2: Installation Qualification of the BLItz System ............................................................................................ 6

2.1. Purpose ...............................................................................................................................................................6

2.2. Materials Required to Install the BLItz System ...................................................................................................6

2.3. System Owner .....................................................................................................................................................6

2.4. Receiving Inspection ...........................................................................................................................................6

2.5. Comments (page intentionally left blank) ..........................................................................................................9

2.6. Setting up the BLItz System ..............................................................................................................................10

2.7. Communication Verification .............................................................................................................................12

2.8. Software Verification ........................................................................................................................................12

2.9. Documentation .................................................................................................................................................12

2.10. Comments (page intentionally left blank) ........................................................................................................13

2.11. Installation Qualification Sign Off ..................................................................................................................... 14

SECTION 3: Operation Qualification of the BLItz System ............................................................................................ 15

3.1. Purpose .............................................................................................................................................................15

3.2. Method .............................................................................................................................................................15

3.3. Materials Required For Operation Qualification ..............................................................................................15

3.4. Quantitation Qualification ................................................................................................................................16

3.4.2. Sample Preparation .................................................................................................................................17

3.4.3. Data Acquisition (Experiment Set Up and Run) Protocol .........................................................................17

3.4.4. Data Analysis Protocol .............................................................................................................................21

3.4.5. Repeat Quantitation Qualification ...........................................................................................................22

3.5. Kinetics Qualification ........................................................................................................................................23

3.5.1. Overview of the Method ..........................................................................................................................23

3.5.2. Sample Preparation .................................................................................................................................23

3.5.3. Data Acquisition (Experiment Set Up and Run) Protocol .........................................................................24

3.5.4. Data Analysis Protocol .............................................................................................................................29

3.5.5. Repeat Kinetics Qualification ...................................................................................................................29

3.6. Comments (page intentionally left blank) ........................................................................................................30

3.6. Operation Qualification Sign Off ....................................................................................................................... 31

SECTION 1: QUALIFICATION DESCRIPTION FOR THE BLITZ SYSTEM

The BLItz system IQOQ user manual includes three sections:

SECTION 1: QUALIFICATION DESCRIPTION DESCRIPTION OF THE OVERALL VALIDATION PROCEDURE

SECTION 2: INSTALLATION DOCUMENTATION AND VERIFICATION OF PROPER

1.4. BLITZ SYSTEM OVERVIEW

The BLItz system user interface is based on five Experiment types:

• Quick Yes/No: tests samples for the presence of a protein.

1.5. COMMENTS (PAGE INTENTIONALLY LEFT BLANK)

AUTHORIZED CUSTOMER REPRESENTATIVE

AUTHORIZED CUSTOMER REPRESENTATIVE

AUTHORIZED CUSTOMER REPRESENTATIVE

SECTION 2: INSTALLATION QUALIFICATION OF THE BLITZ SYSTEM

2.2. MATERIALS REQUIRED TO INSTALL THE BLITZ SYSTEM

• 2.5 GHz Dual Core CPU

• 32-bit or 64-bit Windows XP and Windows 7

• One (1) USB 2.0 port

• One (1) monitor (minimum 1,024 X 768)

2.3. SYSTEM OWNER

2.4. RECEIVING INSPECTION

1. Unpack the box.

TABLE 2.1 BLITZ SYSTEM PACKAGE CHECKLIST

ITEM CHECK IF RECEIVED

BLITZ PRO SOFTWARE CD

BLITZ STARTER KIT