Bio-rad Profinia™ Protein Purification Instrument Manuale Utente

Profinia™Protein Purification SystemInstruction ManualFor Technical Service Call Your Local Bio-Rad Office or in the U.S. Call 1-800-4BIORAD (1-800-42

The following accessories and components can be ordered separately for use with theProfinia system:• Profinia control lysate — dual-tag (histidine and

Warning: Most USB portable memory devices display a flashing light when data is beingtransferred. Wait until the light stops flashing, an indication t

• Calibrate Pressure Sensor9.2.1 Calibrating Touch ScreenTo calibrate the system touch screen:1. Touch the Data/Utilities button from the system hom

5. Touch the Start button; the system pumps water from the water bottle to the waste outlet to remove air from the system and the delivery tube. When

4. Fill a 50 ml sample tube with 45 ml of deionized water, and reserve it for later use (towash sample line 1 when calibration is complete).5. Fill

3. Fill the system’s water bottle with at least 500 ml of deionized water. Empty the wastebottle if needed.4. Touch the Data/Utilities button on the s

To calibrate the pH probe:1. Touch the Data/Utilities button on the system home screen, then touch CalibrationFunctions.2. Touch the up and down arr

• Clean All Inlet and Outlet Lines• Clean Pump Check Valves• System NaOH Wash• Clean Inline Check Valve• Affinity Cartridge Cleaning in Place• Desalti

Fig. 9.5. Diagram of all internal Profinia system hardware components.The adjustable components are:Pump flow rate — set in ml/min. After a flow rate

Outlet select valve (OSV) — rotary valve (10 port) that directs system output. After a valveposition is entered in the Edit menu and you touch the Upd

Manual control toolbar — buttons (5) at the bottom of the Manual Operations screen(Figure 9.6) that control:• Toggle lamp — switches the UV lamp on or

1.4 Related Consumables and ProductsThe following related products are available from Bio-Rad (see Appendix B for a list ofordering information and c

Manual control screen — center section of the screen, for manually operating systemcomponentsToolbar — lower section of the screen, includes command b

9.3.3 Setting Time and DateTo change the system date and time settings (Figure 9.7):1. Touch the Data/Utilities button on the system home screen.2.

9.3.4 Cleaning Sample LinesThis procedure is used to clean the two sample lines; buffer position 8 holds the 20%ethanol solution and is used as a fin

9.3.5 Cleaning Fraction LinesThis procedure is used to clean the eight fraction lines; buffer position 8 holds the 20%ethanol solution and is used as

9.3.6 Cleaning Buffer LinesThis procedure is used to clean buffer lines 1 through 7; buffer position 8 holds the 20%ethanol solution and is used as a

9.3.7 Washing System With EthanolThis procedure is used to clean the complete system with ethanol; all buffer lines, sample lines, fraction lines, ca

9.3.8 Cleaning Inlet and Outlet LinesThis procedure cleans all buffer and sample inlet lines and fraction outlet lines, followed bya system purge wit

9.3.9 Cleaning Pump Check Valves Use this procedure to pump 0.5 M sodium hydroxide (NaOH) solution through the pump toclean the check valves: 1. Tou

9.3.10 Washing System With NaOH Use this procedure to wash the internal system lines (including the cartridge lines) withNaOH solution:1. Touch the

9.3.11 Cleaning Inline Check ValveUse this procedure to manually clean the inline check valve positioned between the cartridge 2 outlet and the fract

Section 2Installation Procedures2.1 System InstallationThe Profinia™purification system is shipped in three boxes. The largest box contains theinstru

3. On the Diag/Maint Functions screen, touch the up and down arrows to scroll throughthe list of functions and highlight “Affinity Cartridge Cleaning

Fig. 9.17. Desalting Cartridge Cleaning in Place screen.Table 9.2 Steps for Desalting Cartridge Cleaning in PlaceFlow rate (ml/min)1 ml Cartridge 5 m

5. Select the OK button to confirm selections and return to the Diag/Maint Functionsscreen.Fig. 9.18. Select End of Sample/Reagent Detection screen.9

Fig. 9.19. Select Method Temperature screen.9.3.16 Select Display ContrastFollow the on-screen instructions to increase or decrease the contrast.Fig.

Section 10 Profinia™Purification Kits10.1 Introduction10.1.1 Background The Profinia protein purification system includes a complete line of consuma

Table 10.1. Profinia purification and buffer kits.Kit Catalog # Solutions Powdered Reagents CartridgesPurification Desalting Storage Buffers Buffer

Native IMAC buffers* are used for proteins that partition into the soluble fraction of bacteriallysates and that have accessible His sequence tags. Ta

Figure 10.1. shows proper placement of native IMAC buffers in the Profinia instrument.Fig. 10.1. Native IMAC buffers properly positioned with instrume

Table 10.3. Formulations for buffers and solutions provided in the Profinia denaturing IMACkits.Profinia Supplied Concentrated Working 1x Supplied V

Figure 10.2 shows proper placement of denaturing IMAC buffers in the Profinia instrument.Fig. 10.2. Denaturing IMAC buffers properly positioned with i

Fig. 2.2. Three boxes containing Profinia instrument and components. 5. Use the handles cut into the side of the top section of the box to lift it of

Table 10.4. Formulations for buffers and solutions provided in the GST kits.Profinia Supplied Concentrated Working 1x Vol, Buffer Solution Formulat

Note: Desalting buffers are already included in native IMAC and GST purification andbuffer kits. Optimized desalting and storage solutions for unique

Section 11 Connecting External ComponentsFunctionality of the Profinia™protein purification system can be enhanced by add-on components. The instrumen

5. Lift off the access panel by pulling it straight toward you (Figure 11.2). Fig. 11.2. Rear view of instrument with access panel removed.6. Locate t

9. Connect the fitting you disconnected in step 8 to the inlet of the pH flow cell. 10. Find the length of tubing supplied with the pH monitor kit. Co

Fig. 11.5a. Installing the flow cell. Fig. 11.5b. Connecting the electrical connector.Fig. 11.5c. Connecting the tubing. Fig. 11.5d. Installing the pH

Fig. 11.6. Installing 37)( tubing into the position 2 buffer line.5. Place the other end of the tubing into the external buffer bottle.Note: If conne

Once the longer tubes have been installed and other standard setup procedures are complete, the system is ready for a purification run.11.2.3 Externa

Fig. 11.8. Profinia cooling accessory. The instrument comes with two cooling units as shown.To install the cooling accessory on the instrument:1. Fil

11.4 Connecting non-Bio-Rad ColumnsColumns from other suppliers can be connected to the Profinia instrument. Figure 11.10displays other commercially

Fig. 2.4. Using lift handles to remove the instrument from packaging.8. Connect the power cable to the back of the instrument (see Figure 2.5 for loc

Section 12 The Profinia SoftwareThe Profinia software runs on a PC platform, and allows viewing and reporting of run data;it is an optional accessory

Fig. 12.1. Profinia software main screen. Area 1, menu options; area 2, directory menu; area 3, file menu;area 4, chromatogram window(s); area 5, data

4. Highlight the directory in your computer to which you’d like the run data transferred.5. Start the purification run according to the procedure ou

12.4.2 Chromatogram OverviewThe following run parameters are displayed in the software chromatogram (Figure 12.3):• UV-1 — data trace from the UV-1 d

5. Repeat this procedure for each data category for which you’d like to implement a colorchange. When finished, click Save to retain changes and retu

Changing Major Steps DisplayedMajor steps are regions between main steps such as sample load begin/end and wash-1begin/end. The following major steps

12.5 ReportsProfinia software enables users to create and print reports for use in presentations or labo-ratory recordkeeping.12.5.1 Standard Report

5. Type a name for the report into the “Name your custom report” text line.6. Click the Save button to save the customized report version. The name

Section 13 Care and MaintenanceProper care and maintenance of Profinia™system parts help keep the instrument operatingat optimum performance levels.13

4. If system power will be left on, go to the Manual Operation screen (Data/Utilities >Diag/Maint Functions > Manual Operation) and turn the la

2.2 First-Time Use of the Profinia SystemBefore using the Profinia system, perform buffer, sample, and fraction line cleaning procedures to flush the

5. Place the cleaning tray in the fraction collection area.6. Touch the Start button. A progress bar displays, tracking the duration of the procedur

13.2 MaintenanceRegular maintenance helps keep the system performing optimally. Figure 13.1 is a diagramof the instrument’s main parts and connection

Use the following procedure to access all internal components, then locate the part-specificprocedure for servicing or replacing the hardware in the s

Diluent bottle — filled with deionized waterWrenches:• Open end 7/16"• Open end 1/2" • Open end 9/16"Replacement check valves (if requi

Cleaning and Replacing Inlet (Bottom) Check ValvesFig. 13.4. Diagram of the inlet check valve.To clean or replace the inlet check valve:1. Remove the

Cleaning and Replacing Outlet (Top) Check ValvesFig. 13.5. Diagram of the outlet check valve.To clean or replace the outlet check valve:1. Remove the

Note: Do not overtighten the fittings. Finishing Pump Check Valve Maintenance ProceduresWhen finished with check valve cleaning or replacement procedu

Fig. 13.6. Removing the inlet tube from the pump assembly.4. Use your fingers to remove the 1/16" outlet tube from the front of the pump body; y

Fig. 13.8. Pulling the pump head away from the drive assembly. 7. Using your fingers, remove the pump wash fittings from the top of the pump head. 8.

12. Press the new seals into the recesses in the pump body with your fingers, then use thepiston seal tool to snap them into place. Soak the pump body

4. Fill the four 125 ml and three of the 250 ml buffer bottles provided with the instrument tocapacity with deionized water, and place them in buffer

Fig. 13.12. Align the flat side of the guides with the carrier assembly recess (tubing removed for clarity).18. Use the 9/64" Allen wrench to tig

the UV monitor cuvette. If the reference voltage is less than 0.05 volts, the lamp intensity islow and the lamp should be replaced. If the sample volt

Fig. 13.13. UV monitor assembly.3. Follow the lamp’s power cable to the metal panel, and unplug it (Figure 13.14). Note: Depress the locking tab befo

Fig. 13.15. Removing UV lamp and filter carrier.5. Using the 5/64" Allen wrench provided with the new lamp, remove the set screw holdingthe UV l

Fig. 13.17. Sliding the replacement UV lamp out of the carrier. 7. Remove the new UV lamp from its package. Note the round window for the UV light(F

Fig. 13.19. Filter carrier and UV lamp carrier.11. Place the lamp and filter carriers on the UV module body and secure with the screwsusing the 9/64&q

Fig. 13.20. Removing the two tubing fittings from the top of the flow cells; follow this procedure for eachflow cell.4. Using a 9/64" Allen wren

Fig. 13.22. Removing one of two screws holding the flow cell assembly in place.8. Gently pull the flow cell assembly from its recess.9. Look through

14. Turn on the unit and wait 5 min.15. Check the sample and reference voltages, as described above. 13.2.5 Buffer and Sample Inlet TubesIf the buffe

Table 13.1. Diameters and lengths required for Profinia system tubing connections.System Connection System Label Tubing OD, Tubing Inches Length,Inche

It is recommended that this function be performed a second time prior to first use to removeany air that might have entered the system. The system has

System Connection System Label Tubing OD, Tubing Inches Length,InchesOutlet select valve port 10 to waste manifold OSV P10 to waste 1/8 8Column valve

Fig. 13.24. Schematic diagram of the Profinia system.To install the replacement tube:1. Remove the damaged tube and measure it, or refer to the list

5. Slip the lock ring over the tubing. Note: The lock ring is tapered on the inside; the wide end of the lock ring faces the cut endof the tubing.6.

13.2.7 Inline FilterThe inline filter is a 6 µm polyester membrane filter, which protects the system from particlesin the inlet fluid stream. The inl

13.2.8 Power ConditioningThe Profinia system is designed to operate in a laboratory environment with other instrumentsnearby; it complies with the CE

Appendix AProfinia™System Specifications Operating limitsFlow rate range 0.2–20 ml/minMaximum back pressure 45 psi (3.4 bar) Detection UV detection

Appendix BProfinia™System Ordering InformationCatalog # Description156-3004 Profinity eXact Antibody Reagent620-0010 Profinia Software, includes USB c

Catalog # Description620-0226 Profinia GST Purification Kit, 1 ml, includes Profinia GST buffer kit, 2 x 1 ml GST and 2 x 10 ml desalting cartridges62

Catalog # Description620-0415 Profinia Instrument Tubing Kit, includes assembled system tubing andfittings 620-0416 Profinia UV Flow Cell Replacement

Appendix CProfinia™System TroubleshootingProblems and SolutionsProblem SolutionNo power to system If the instrument will not start, check that the sys

3. Place the tube in the sample 1 position and select the “1 Sample” option in the appropriaterun set-up screen.4. Prepare the buffers and reagents

Problem SolutionIf there aren’t problems noted with sample application, usethe “Check Flow Rate” utility (section 9.2.2) to check if theflow rate is c

Problem SolutionData does not display The Profinia system must be connected to the computer via on a computer USB cable, and Profinia software must b

Instrument Error Codes and SolutionsERROR ERROR MESSAGES Possible Cause(s) and CODE Solution(s)2 CANNOT TALK TO I2C DEVICE XX. Inter-processor commu

Appendix DProfinia™System Error MessagesThe error messages listed below are displayed on the Profinia touch screen when instrumentoperation errors occ

Appendix ESample PreparationPreparing Lysates Prior to PurificationLysates from E. colicultures can be prepared using conventional sonication procedur

Native Lysate Preparation Using Profinia Bacterial Lysis/Extraction Reagent (Recommended for IMAC procedures; binding capacities of GST fusion protei

Appendix FSample Preparation, Application, and Analysis TipsTopic Tips and RecommendationsSample PreparationSample resuspension Resuspend harvested ba

Topic Tips and RecommendationsSample Screening — Before choosing a native or denaturing purification protocol, Solubility Profile approximate express

Topic Tips and RecommendationsSample Application Load and Flow Rate The yield of the target protein is dependent on the flow rate at Dependency which

Topic Tips and RecommendationsPrecipitation of Precipitation of the elution peak can occur when the eluted protein Elution Peak is too highly concent

Fig. 2.11. Typical Experion™system electropherogram of the purified fraction of the native IMAC controllysate purification. Expected Results for GST P

Topic Tips and RecommendationsFor longer-term storage, any unused buffer can be stored at–20°C and used within 6 months. Upon thawing, check for anypa

Appendix GProfinia™System Annual MaintenanceThe list of maintenance procedures in Table G should be performed regularly to ensureoptimum instrument pe

Date Maintenance Procedure Performed Comments4. Replace the UV lamp, if necessary (section 13.2.3).5. Check UV flow cell assembly (section 13.2.4).Aft

Appendix HProfinia™ Instrument Cleaning and Maintenance Log User Information Cleaning Maintenance Miscellaneous Date Operator Short Term Long Term

Appendix IProfinia Protein A and G Method1.1 Background InformationThe Profinia protein purification system is an automated, user-friendly instrument

Table 1. Protocol steps for Profinia 1 ml protein A plus 10 ml desalting method with asingle sample1.3 Required Equipment and Materials Bio-Rad Affi-

In order to preserve the activity of labile antibodies when using acidic elution conditions, it isrecommended that 1 M Tris-HCl, pH 8 be added to the

Section 2Guidelines When Using Protein A Methods 2.1 Sample Preparation InformationAffi-Prep protein A cartridges are convenient, ready-to-use device

For additional cleaning-in-place options for protein A cartridges, refer to the Utilities sectionof the manual for information on the affinity cartrid

Fig. 1. Typical chromatogram and SDS-PAGE for Bio-Rad 1 ml Protein A plus 10 ml desalting method.Section 3Frequently Asked Questions and Troubleshooti

Contact Bio-RadBio-Rad Laboratories, Inc.2000 Alfred Nobel DriveHercules, CA 94547Phone: (510) 741-1000Fax: (510) 741-5811Bio-Rad Technical Support T

Fig. 2.13. Typical gel result of the GST control lysate purification. Lane 1, Precision Plus™MW standards;2, skip; 3, load; 4, flow-through; 5, wash;

Section 4Ordering InformationCatalog # Description732-0111 Luer to M6 Adaptor Fittings Kit732-0112 Luer to 10-32 Adaptor Fitting Kit732-0113 Luer to B

Appendix JProfinia Profinity eXact Method ManualSection 11.1 Background InformationThe Profinia protein purification system is an automated, user-fri

Table 1. Method Steps for Bio-Rad Profinia 1 ml eXact plus 10 ml Desalting Methodwith 1 Sample1.3 Required Equipment and Materials Bio-Rad Profinity

Table 2. Recommended Formulations for Buffers and Solutions for 1 ml eXact + 10 mlDesalting Method*These buffers and solutions are available as part

Table 4. Buffer and Chemical Compatibilities for the Profinity eXact Tag System1.4 Recommended Storage ConditionsPurchased buffers obtained from Pro

and binds to the N-terminal co-expressed affinity tag in the fusion protein. Subsequent to thecolumn binding and washing steps, the eXact protease per

For additional cleaning-in-place options for eXact affinity and desalting cartridges, refer tothe Utilities section of the manual for information on t

Fig. 1. Typical Chromatogram and SDS Page for Bio-Rad 1 ml eXact plus 10 ml Desalting Method*.Profinity eXact affinity-tagged maltose binding protein

Low purity of target protein Sample load and wash steps Select two wash option for eXacton SDS-PAGE or residual do not return to baseline with meth

Appendix KProfinia Program Methods Planning WorksheetsThe worksheets included in this section are designed to help plan desired changes to stepparamet

Section 3Key Terms and Definitions 3.1 Bio-Rad MethodsThe following automated methods — Bio-Rad methods — for common affinity-tagged protein purifica

203

204

205

206

207

208

209

210Method Type:Method Name:eXact Cartridge Size:# Cartridges:Sample 1 Name & Volume:Sample 2 Name & Volume:Method Step Step DescriptionPositio

211Method Type:Method Name:eXact Cartridge Size:Desalting Cartridge Size:Sample 1 Name & Volume:Sample 2 Name & Volume:Method Step Step Descri

212

3.3 Toolbar Navigation ButtonsToolbar navigation buttons are located at the bottom of each touch screen (Figure 3.1).Fig. 3.1. Toolbar navigation but

213

214

TrademarksPentium is a trademark of Intel Corporation. Windows and Windows XP are trademark are trademarks of Microsoft Corporation.215

Life ScienceGroup00-0000 0000 Sig 0308Bulletin 0000 Rev A US/EGBio-Rad Laboratories, Inc.Web site www.bio-rad.com USA 800 4BIORAD Australi

• Conc. — buffer position in the instrument and buffer concentration (the instrumentdilutes concentrated buffers)• ml/min — flow rate displayed in ml/

3.5 Alphanumeric and Numeric Keypads3.5.1 Alphanumeric KeypadText is entered into the alphanumeric keypad via a text line at the top of the screen (

Fig. 3.4. Numeric keypad.18

Section 4Profinia System Quick Guides4.1 Unpacking and Installing the Profinia Protein Purification System19

20

21

Installing the Profinia Software22

Caution!Please read all safety-related information before operating the Profinia system. This instrument is for laboratory use only.Always connect the

4.2 Profinia™System Quick Guide23Note: An “X” indicates that no reagentor container is required in that position.Place bottlecaps hereduring the runB

242B. Waste and Water Bottles1. Empty the waste container, and check the diluentwater bottle for sufficient volume.2C. Sample Tube Installation1. Add

25Step 3: Cartridge InstallationIf cartridges are not already installed, touch the NO button. You will proceed to the Setup CartridgePositions screen.

265. Remove the top cap from the cartridge. Move the uppercartridge holder down to the cartridge.6. Twist the black collar to tighten the upper fittin

Section 5Home Screen OverviewThe LCD touch screen on the front of the instrument is used to control Profinia™systemfunctions (Figure 5.1). You may use

Warning: Overwritten run data files can no longer be retrieved; the data is permanentlydeleted. • Calibration — system functions enable calibration o

Section 6Bio-Rad MethodsThe Bio-Rad Methods function of the Profinia™ system is used to select preprogrammedpurification and desalting methods. These

Fig. 6.1. Select Method Temperature screen.3. Touch the arrow button to select “4°C” or “Room Temperature”, then touch the OK button to return to the

Figure 6.2 shows the screen sequence when starting a Bio-Rad method.Fig. 6.2. Screen sequence when starting a Bio-Rad method.31

To begin a Bio-Rad method purification procedure, touch the Bio-Rad Methods button inthe lower toolbar of the home screen (Figure 6.3). You will proce

Table of ContentsPageSection 1 Profinia Protein Purification System Description ...11.1 Profinia System Overview...

Fig. 6.4. Select Method system. Fig. 6.5. Select Method Type & Options screen.To select the method type and options: 1. Touch the up and down arr

Note: Two samples can be run for all methods except the desalting-only method. Whentwo samples are selected for affinity-only methods, the number of c

The Bio-Rad methods function provides two sample loading flow rate options:• Standard — recommended for normal- to high-expression proteins (<100 k

Elution of target protein is typically conducted by incubating the resin in the appropriate bufferat room temperature (see Appendix J for buffer formu

Fig. 6.8. Enter Run & Sample Information screen.Note: When two samples are selected for the method, press the View Method button in thelower toolb

• A280of 1 mg/ml — extinction coefficient for the protein expressed as the absorbance ofa 1 mg/ml solution of the protein of interest at 280 nm for a

Fig. 6.9. Edit Lot Number Information screen.3. Touch the arrow buttons to the right of the lot number information text box to scrollthrough the list

Fig. 6.10. Install Reagents, Sample & Fraction Tubes screen (left) with arrows demonstrating the correlationbetween the diagram and placement of r

• Fraction collection tubes — indicated on the touch screen by a set of four tubes onthe left labeled 1A through 1D for sample 1 collection, and four

Fig. 6.12. Replace buffer caps with buffer lids before installation.4. Open the lid on the Profinia instrument to access buffer sipper tubes and the

6.7.1 Viewing Steps...556.7.2 Viewing Real-Time Data With Profinia Softwar

Important: It is important to orient the buffer bottles properly in the instrument; the graduatedmarks on the buffer bottles are used to determine how

Note: Some kits do not require a reagent bottle in every position. If a position does notrequire a bottle (indicated by an “X” on the installation scr

6.5.3 Installing Fraction Collection TubesThe fraction collection area is at the bottom of the Profinia instrument (Figure 6.17).Fractions are collec

Fig. 6.18. Installation of a tube into fraction tube position 1D. When all fraction tubes required in the run have been installed, touch the Next butt

Fig. 6.19. Select cartridge status screen.6.6.2 Setting Up Cartridge Positions for PrimingFollow the instructions on the screen to prepare both cartr

6.6.3 Priming the Cartridge LinesThe Priming in Progress screen (Figure 6.21) displays a progress bar that indicates thepercentage of priming time co

Fig. 6.22. Install Bio-Scale Mini Cartridges screen.To install new cartridges:1. Disconnect the upper and lower cartridge fittings (Figure 6.23).Fig.

Fig. 6.24. Inserting the cartridge into the cartridge 1 position; twist the cartridge in a clockwise directionas indicated by the arrow to fit the bot

Fig. 6.25. Completing the cartridge installation procedure.6. Repeat this procedure to install a Bio-Scale Mini cartridge into the cartridge 2 positi

Fig. 6.26. Lower fitting tightened into place (left) and upper cartridge fitting tightened into place (right);cartridge 2 position remains closed.When

9.1.2 Real-Time Data Transfer to Profinia Software...939.2 System Utilities ...

Fig. 6.27. Install Bio-Scale Mini Cartridges screen showing sample loop installation in the cartridge 1position.To connect and fill the desalting loop

4. To apply sample, fill the syringe and insert it into the open female luer of the 3-waystopcock. Turn the 3 way stopcock lever up (toward the top i

Fig. 6.30. Start Run screen. 6.7.1 Viewing StepsTouch the View Steps button (available on the Start Run screen at any point during the run)to display

Fig. 6.31. View Method Steps screen. 6.7.2 Viewing Real-Time Data with Profinia SoftwareThe Profinia system does not require software and a computer

2. Open Profinia software on your computer by double-clicking on the desktop Profiniasoftware icon. Look for the “Waiting for data” message in the lo

Once memory has cleared, the method starts and the Run in Progress screen displays.6.8 Monitoring Run Progress6.8.1 Run in Progress ScreenThe Run in

The lower part of the Run in Progress screen shows the real-time data:• Real-time data at — system clock time. The date and time setting is available

Note: The peak detect delay and max peak volume parameters can only be edited in theprogram methods mode (see section 7.3.1).• Peak detection sensitiv

Fig. 6.36. Touch the Manual Peak Divert button at the step in the run indicated by the arrow to optimizeconcentration of the eluted protein.When the p

6.8.4 Canceling or Pausing a RunTo cancel the run (when you cancel a run, you proceed to the Prepare System to CleanLines screen):1. Touch the Cance

12.4 Viewing Run Data Files...13512.4.1 Opening Files in Profinia Software ...

2. Place tubes containing 10 ml of deionized water in both sample 1 and sample 2 positions;place water in both sample positions even if only one samp

Fig. 6.40. Run Completed screen. 6.10.1 Exporting Data to a ComputerThe run data file can be transferred to a portable memory device as an .ofi file,

Re-Running a MethodTo repeat the method just performed with the same buffers, touch the Re-Run Method button in the toolbar at the bottom of the End o

Fig. 6.42. Prepare System to Clean Lines, short-term wash selected screen.To prepare the system for the short-term wash, follow the onscreen instructi

Fig. 6.43. Prepare System to Clean Lines, long-term wash screen.To prepare the system for the long-term wash:1. Place bottles of deionized water, 20%

Section 7Program MethodsThe program methods function of the Profinia™ system is used to create methods by editingthe Bio-Rad purification and desaltin

Fig. 7.1. Screen sequence when starting a program method run. To create a method, touch the Program Methods button in the lower toolbar of the homescr

Fig. 7.2. Select Method System screen.7.2 Selecting Method Type and OptionsThe Select Method Type & Options screen (Figure 7.3) allows you to ch

To select the method type and options:1. Touch the up and down arrows to select the method template from the list of optionsdisplayed on the screen.N

Fig. 7.4. Enter Run & Sample Information screen. The following method-specific information can be entered or selected:• Method name — name for th

Section 1Profinia™Protein Purification System Description1.1 Profinia System OverviewThe Profinia protein purification system is an automated chromat

• Volume — user-entered sample volume between 2 and 999 ml. If you have chosen toperform a run with two samples, a volume must be entered for each sam

3. Touch the arrow buttons to the right of the lot number information text box to scrollthrough the list of buffers, solutions, and kit information r

Fig. 7.5. Method Information screen. The edit button displays only if the highlighted step has editable parameters.The View Sample button appears for

Most steps show the following information and parameters:• Method name — editable in the Enter Run & Sample Information screen• Step name — cannot

Fig. 7.7. Select Step Parameter to Edit screen with parameters for protein elution steps displayed.The peak detection parameters that can be edited f

When all edits are completed, touch the Save button in the Enter Run & SampleInformation screen to store your method in system memory. As your met

• Waste bottle — left side of the touch screen and positioned at the lower left of theinstrument. On the touch screen, the waste bottle is displayed w

Note: Reagent volumes that exceed system container sizes can be used in external containers connected to the instrument via extra tubing. Make sure to

• Install Cartridges — displays the required cartridges and their respective positionsWhen cartridge installation is complete, touch the Next button t

Fig. 7.11. Start Run screen directs application of the sample to the sample loop (desalting-only method).4. To apply sample, fill the syringe and ins

2• Built-in cleaning and storage steps — automated system cleaning ensures continuedpeak instrument performance• Purification kits — affinity and desa

7.7 Viewing Real-Time Data With Profinia Software To collect real-time data:1. Connect the USB cable provided with the Profinia software package to

7.8.1 Viewing StepsTouch the View Steps button (available on the Start Run screen, and at any point during therun) to retrieve the View Method Steps

3. Touch the Prime button again to immediately start priming the next selected reagentline upon completion of the first selected line.4. Repeat unti

7.9.2 Collecting Purified SampleThe purified sample is collected in fraction tube position 1D or 2D. When the purified protein is collected, the Run

Fig. 7.16. Run Completed screen.7.11.1 Exporting Data to a ComputerThe run data file can be transferred to a portable memory device as an .ofi file,

Important: When the Profinia system is used in a cold environment, it is best to leaveinstrument power on; the slight amount of heat generated helps c

Section 8 Saved MethodsThe Profinia™ instrument can store up to 35 program methods in memory. Saved methodscan be retrieved alphabetically according t

2. Use the arrows to select the viewing mode by: “Method System” or “All MethodSystems”. Touch the Next button to proceed to the Select Saved Method

screen (Figure 8.3). See section 7.3 for details on editing the method and entering run andsample information.Fig. 8.3. Enter Run & Sample Informa

Section 9 Data and UtilitiesThe Data/Utilities screen on the Profinia™purification system provides access to maintenance, diagnostic, and calibration