Proteomics PA 800 (Beckman) ProteomeLab System (Detect Instrument Picture to Jump to that section of SOP)

Transcription

1 Beckman PA800 Proteomics PA 800 (Beckman) ProteomeLab System (Detect Instrument Picture to Jump to that section of SOP) Windows Login: Username: Administrator Password: gold To Instrument 32 Karat sw: Proteanlab/pa800 Instructor/15pa800 Student/student Prepared by: Bob Morrison STLCC-CPLS, Instrumentation Specialist Dec 08, rev Mar2012 Page 1

2 Proteomics: PA800, Overview, Specifications The PA 800 plus Pharmaceutical Analysis System provides a comprehensive, automated, and quantitative solution for the characterization and analysis of proteins. Innovative system design ensures dependable operation and durability. The PA 800 plus application menu includes SDS-gel molecular weight analysis, charge heterogeneity analysis utilizing advanced capillary isoelectric focusing technology, and carbohydrate profiling for the assessment of glycoprotein microheterogeneity. PA800, PN , SN , Beckman SysID: , Delivered 1/10/12, Installed at BRDG 2/13/12 Hardware setup training: 2/24/12 with Paul Flynn to RH, BM Software/Application training: TBD idth: 25 in (63.5 cm) Height: 29.2 in (74.2 cm) Height with Door Open: 38.8 in (98.6 cm) Depth: 28.4 in (72.1 cm) Weight: 188 lbs 85.3 kg) includes photodiode array detection lectrical Requirements: Voltage V 50/60 Hz uffer Tray: 2 x 36 universal vials Current Range: 3 to 300 μa programmable at 0.1 μa Increments etectors: Diode Array, UV/Vis, aser-induced Fluorescence Pressure Range: -5 to +100 psig (Rinse/Separations), -5 to +25 psig (Sample Introduction) ample Temperature Control: 4 C - 60 C apillary Temperature Control: 15 C - 60 C ample Tray: 2 x 96-well plates, 2 x 48 2 ml vials, 2 x ml vials, 2 x 48 PCR vials oltage Range: 1 to 30 kv programmable at 0.1 kv Increments NSPSC: Mercury Content: Item Contains Mercury - Dispose According to State/Local Law Page 2

3 Protoemics: ProteomeLab, Plans, Info, Manuals Capillary Electrophoresis Beckman ProteomeLab PA800 ( Rafael Hernandez 2/28/12) This is a protein characterization system used intensively today by biopharmaceutical companies such as Pfizer, Johnson and Johnson, and many others. This instrument is capable of performing identity, purity and heterogeneity tests on nano-liters amount of bio-therapeutics drugs. The PA800 also has automated capabilities allowing us to test multiple samples in a very short period of time. The ProteomeLab PA800 will be introduced to our students in the Biotechnology Program enrolled in classes such as Bio219, Bio220 and possibly during Advanced Topics Courses (Bio226). The student will learn the theory, applications and operation of the system. Hotlink to Beckman Capillary Electrophoresis Basics Presentation.. Pdf Hotlink to Beckman Capillary Electrophoresis of Proteins Presentation.. Pdf Hotlink to Beckman IgG Assay Data Sheet.. Pdf Hotlink to Beckman Peptide Mapping Brochure.. Pdf Hotlink to Beckman PA 800 Users Guide.. Pdf Page 3

4 Protoemics: ProteomeLab, PA 800 Related Videos (found by Betsy Boedeker Sep 2012) The BC ProteomeLab PA 800 System Part 1 Part 2: Further Inside the Front Cover & Removing the Spring Injectors Part 3: The System Light Indicators & Inside the Bottom Door Replacing the Ejector and Electrode on the PA 800 plus and PA 800 Enhanced Installing a Capillary Cartridge Removal Capillary Storage Page 4

5 Proteomics: PA800, Hardware Diagram Instrument on/off Page 5

6 Proteomics: PA800 Startup and Admin 1. Turn on PA-800 (front button) before starting PC 2. Boot PC and login windows as usersname: Administrator, password: gold 3. Start 32 Karat software application and note configuration of instrument from available icons: CHO (not loaded), IEF, IgG purirty, SDSMW, Performance 4. Choose Performance, Select Project and login as username: ProteanLab password: pa Assign users to projects with appropriate privileges. Select Tools, Select System Wizard, Select Project and assign users to each, moving most privileges to all but generally recommended to remove some such as options to delete or purge system log files Repeat until all active projects (SDS, Performance, Peptide, IgG) are loaded. Note CHO and Default are not active Page 6

7 Proteomics: PA800, Tray Handling System Sample Inlet (SI) (temp regulated) Sample Outlet (SO) Buffer Inlet (BI) (not temp regulated) Buffer Outlet (BO) Note Plate Orientation: ABCD readable from front Page 7

8 Proteomics: PA-800, Vials and Supplies Highly recommended that all vials be ordered thru Beckman and/or National Scientific to avoid quality issues Target Vials, National Sci, C4010-1B6 PCR holder vials PN ; holds standard PCR type tube inside, spring loaded, with gray septa/cap required Gray cap for PCR vial holders; PN Recommended that vial caps be loaded with slight twisting motion as you press down. Do not crimp or block semi-circle opening in caps Page 8

9 Proteomics: PA800, Capillary Cartridge Construction Inlet 1. Supplies; locate Capillary Cartridge PN AE with PN , 691, 692, 693, inside 2. Apertures: PN , Capillary filament : PN Measure to 60.2 cm and cut from longer line 5. Insert line at Outlet side, thread through carefully pushing into plastic tube (1) and Stop when tiny 1cm window is visible in Aperture hole (3) 6. Add seals on ends, then recheck alignment of window in aperture opening (3) 7. Layout cap fitting on black metal template to make cutoffs with square scoring tool provided in kit. 8. Use eyepiece to verify clean cut on both ends with no burrs or rough edges. Sand using other side of square tool if needed, re-examine. 9. Add seals to capillary lines 10. Insert aperture (3) into hole careful not to break or force capillary filament. 11. Insert O-ring from opposite side of fitting to inside of aperture. 12. VERY SOON after this is done put entire Capillary fitting into the case and fill with DI water. Cap/filament ends must be kept wet before, during And after each run. 13. Insert into instrument, make sure capillary lines Are shorter than Electrodes. If not, push filaments higher into plastic tube or recut Outlet Page 9

10 Proteomics: PA800, Misc Notes on Conditioning Run Open Method, select New Method to set conditions for defaults Detector, set a wavelength of 214 Time program; Event-arrow-rinse-pressure (20), time 1.0 min, Forward File-save methods-save as Tutorial (name) Located Capillary performance test kit in 4C, PN Fill vials to 80% level, seal with blue caps Basic Wash,.1N NaOH PN: Running Buffer: PN , 50ml Injection step:.5 psi pressure Separation step; Voltage, normal polarity, 30kv, 6 minutes, Autozero step; 2 min Save Method Whenever you install a new capillary you must make a condition run. Page 10

11 Proteomics: PA800, Page 11

12 Proteomics: PA800, Software from website The PA 800 plus Software includes features that allow you complete control of the PA 800 plus and P/ACE MDQ Systems. The software includes methods and sequence table development capability, advanced data analysis functionality, and includes the analytical tools necessary for proper integration and quantitation of data. In addition, advanced custom reporting can be done in order to output results the way you want to see them. The Direct Control capability allows for more direct user interaction with the system. Additionally, the software also allows for single point control for all detectors configured on these systems. The PA 800 plus software quickly guides users from set-up through routine system operation. Large icons provide intuitive guidance for navigation while on-screen cues indicate system progress at a glance. Insightful Help menus and descriptive system prompts further simplify operator learning, making transfer of PA 800 plus technology to other analysts easier. Ultimately, using PA 800 plus software is as easy as 1, 2, 3. Page 12