发展中的层析技术 - ebiotrade.com avant.pdf · Step elution level Cut OD Elution Additive...
Transcript of 发展中的层析技术 - ebiotrade.com avant.pdf · Step elution level Cut OD Elution Additive...
发展中的层析技术
ÄKTA™avant
技术支持:王济源
15821989818
2 /GE 28-9577-83 /
Version2.0
Evolution of ÄKTA™ 1996 - 2010
1996 - ÄKTAexplorer™
1997 - ÄKTApurifier™
1998 – ÄKTAFPLC™
1999 - ÄKTAprime™
2000 – ÄKTA OligoPilot
2001 – ÄKTA add on´s
2002 – ÄKTA 3D Kit and ÄKTApilot™
2004 - ÄKTAxpress™
2005 – ÄKTAcrossflow™
2006 – New ÄKTApurifier
2009 - ÄKTAmicro™
2009 –ÄKTA25
2010 – ÄKTA150
3 /GE 28-9577-83 /
Version2.0
Customer input Key to product definitionThe definition process for ÄKTAavant™ started in 2005 with a series of customer interviews in North America, Europe and Asia…
… and others
4 /GE 28-9577-83 /
Version2.0
客户的需求
快速: Achieving robust, well characterized processes consumes time. ÄKTA avant overcomes this, enabling quicker insight and allowing you to get more done in the same time frame.
安全: Working at this enhanced pace often results in more errors. ÄKTA avant allows you to obtain reliable results while working in a compliant, error-free manner with complete sample security.
可放大性: ÄKTA™ avant enables you to achieve direct, dependable scalability while systematically anticipating errors to prevent any negative impact on safety, consistency, or economy of the full-scale separation.
5 /GE 28-9577-83 /
Version2.0
ÄKTA™ avant and UNICORN™ 6.0
6 /GE 28-9577-83 /
Version2.0
ÄKTA™ avant设置未来分离纯化的新标准
快速
Inspired security
Inspired scalability
7 /GE 28-9577-83 /
Version2.0
来自“速度”的压力
验证猜想
科研进度
产品上市
解放劳动
8 /GE 28-9577-83 /
Version2.0
ÄKTA™ avant工艺开发的加速器Ultimately it all boils down to getting new products to the market faster. ÄKTA™ avant has the capacity to significantly shorten the process development time.
DoE协助您通过有限的实验次数得到尽可能多的信息
使用BufferPro功能自动化准备缓冲液
模块化的方法编辑快捷方便
• Design of Experiments (DoE) is integrated in UNICORN 6
• Leads the way to automated process development
• Allows you to obtain the maximum amount of information from a minimun number of experiments
• Significantly reduces process development costs
• Completely automated, advanced on-line buffer preparation
• Increases your throughput
• True buffer mixing with corresponding acid and base
• Easy and quick pH-scouting
• Simple, highly intuitive method creation
• Drag-and-drop predefined phases from the Phase Library
• Full programming control still available for the advanced user
9 /GE 28-9577-83 /
Version2.0
Design of ExperimentsDoE 是利用统计学方法对多因素函数合理安排实验点,以减少实验次数得到全面信息,建立可验证的数学模型,用来预测变量对函数的影响。
See film clips on the following slides!
10 /GE 28-9577-83 /
Version2.0
Introduction
Responses:
External data:
Capacity
DBC (Frontal analysis)
Yield
Purity/Selectivity
Molecular weight
Activity
HCP
DNA
Aggregates
Protein A
Peak Data:
Area
Concentration
Amount
Resolution
Asymmetry
Plates per meter
Factors:
Load pH
Load conductivity
Load concentration
Mass load
Wash volume
Wash pH
Wash conductivity
Elution pH
Gradient elution
Step elution level
Cut OD
Elution Additive
Media type
Column size
Bed Height
Flow rate
Residence time
Sample conditions
Wash conditions
Elution conditions
Entire process
Complexity vs. State of the art PD- The possibility to automate process development
11 /GE 28-9577-83 /
Version2.0
Introduction
Process knowledge, development and outcomeTraditional approach: Parameter 1 Fixed, Parameter 2 changed or Parameter 2 Fixed, Parameter 1 changed
DoE arranged approach: Vary all simultaneously
- ”Design of Experience”
Optimum
Parameter 1
Parameter 2
Para
met
er 2
Parameter 1
12 /GE 28-9577-83 /
Version2.0
DoE reduces your costDesign of Experiments allows the maximum amount of information to be obtained from a minimum number of experiments. By improving efficiency, both time and money are saved
• Design of Experiments (DoE) is integrated in UNICORN™ 6
• With DoE, you can reach the same result in 16-32 experiments as you would with 128 experiments using the traditional approach!
DoE Structured approach
Traditional approach
See film clip on the following slide!
13 /GE 28-9577-83 /
Version2.0
ÄKTA™ avant 连接一条高通量工艺开发途径
• Recommended workflowPreDictor™ plates in combination with Assist Software allow screening of a large experimental space toidentify the sub-space that is favorable with respect to one or several defined responses. Once thissubspace has been found, optimization may be performed on columns and ÄKTA™ avant prior to a successful scale up.
• UNICORN™ and Assist are complementaryThe same user interface has been used for UNICORN and Assist in order to facilitate the useof both Softwares.
PreDictor PlatesAssist Software ÄKTA™ avant
UNICORN™6.0
Further optimization & Scale-Up
14 /GE 28-9577-83 /
Version2.0
BufferPro – 自动在线配置缓冲液
ÄKTA™ avant includes BufferPro, truly automated on-line buffer preparation - saves 30 min per buffer and pH-value!
• 种类多Versatility with 26 buffer systems available
• 用相应的酸或碱调pH Buffers are made in analogy with manual preparation: Corresponding acids or bases used for pH adjustment
• 全新设计的Q-阀
• 可以设置浓度Control of conductivity (buffer concentration) for purification sensitive to ionic strength
• Actual mixing ratios and buffer recipes are provided
pH 5.5pH 5.4pH 5.3
16 pH steps 0.1 units apart,with no manual buffer titration at all!
HiScreen™ Capto™ SpH from 4,0 to 5,5 using the CIEX mix buffer
See film clip on the following slide!
15 /GE 28-9577-83 /
Version2.0
方法编辑直观易用
UNICORN 6 includes a new Method Editor, replacing the method wizard. No new text programming is required, new components immediately ready - save a day’s work!
• Contains all instructions used for controlling the chromatographic run
• Easy and flexible method creation - Drag and drop from the phase library into the method outline
• Use pre-defined methods / phases or program your own
• Automatic calculation of volume, pressure and flow rate settings for columns
• Built-in application support
• New GUI for easy viewing and editing of run properties
See film clip on the following slide!
16 /GE 28-9577-83 /
Version2.0
New user interface in UNICORN™UNICORN 6 has an improved graphical interface with a task bar, customizable docking windows, and object navigators that significantly decreases learning time!
UNICORN™ 6
17 /GE 28-9577-83 /
Version2.0
ÄKTA avant硬件设计ÄKTA™ avant sample and system pumps run independently and no re-plumbing is necessary, - save a day’s work per week!
Save time on …• 样品泵
• 上样阀
• 柱位阀
• pH流动池阀
• 压力感受器
System and sample pumps
18 /GE 28-9577-83 /
Version2.0
System pumps
The system pumps are used to generate isocratic or gradient elution in purification methodsFunction
• Isocratic elution• Gradient elution
Details• System pump A and B• Flow rate range 0.001 - 25
ml/min • Maximum operating pressure
20 MPa • Purge valves• Pump piston rinsing system
Pumps
19 /GE 28-9577-83 /
Version2.0
Sample pump
The sample pump is used for direct loading of sample on a column, or to fill sample loops
Function• Pump sample into the flow
path
Details• Flow rate range 0.01 - 25
ml/min • Maximum operating pressure
10 MPa • Purge valves
• Pump piston rinsing system
Pumps
20 /GE 28-9577-83 /
Version2.0
Sample inlet valve
Enables automatic change of different samples
Function• Select different sample
inlets• Integrated air sensor for
injecting all sample• Dedicated buffer inlet
– Fill the sample pump with buffer before sample is introduced
– Wash out sample between runs
Valves
21 /GE 28-9577-83 /
Version2.0
Inlet valves A and BEnable automatic change between different buffers and wash solutions
Function• Change between buffer
and wash solutions• Generate gradients by
mixing buffer A and buffer B
• Integrated air sensorDetails
• 7 buffer inlets (A1-A7, B1-B7)
• 1 buffer inlet (Q) per inlet valve is connected to the quaternary valve
• Transposed position of the inlets
Valves
22 /GE 28-9577-83 /
Version2.0
Injection valve
Enables sample application onto the column
Function• The sample can be injected
using– A syringe into a capillary
loop– The Sample pump into the
sample loop/Superloop™– The Sample pump directly
onto the column• No need for re-plumbing
Details• Flow from all pumps• Sample loop/Superloop • Syringe connection
Valves
23 /GE 28-9577-83 /
Version2.0
Column valve
Enables selection of the column to be used
Function• Integrated pressure sensors
– Column hardware– Column media
• Integrated bypass of columns• Integrated reverse flow
functionality
Details• Up to five columns• Column pressure sensors
– Inlet port– Outlet port
Valves
24 /GE 28-9577-83 /
Version2.0
pH valve
Enables pH measuring and insertion of flow restrictor in the flow path
Function• pH-cell optionally
selectable • Flow restrictor optionally
selectable • Easy calibration of the pH
electrode• Storage of the pH
electrode
D t il
Valves
25 /GE 28-9577-83 /
Version2.0
Outlet valve
Enables collection of large fractions
Function• Outlet fractionation • Directs buffer solution to
the fraction collector
Details• 10 outlet port (Out 1-
Out10)• Waste port• Port to Fraction collector
Valves
26 /GE 28-9577-83 /
Version2.0
Column pressure monitors
Enable measurement of pressure before and after the column
Details• Pre column pressure
monitor (inlet of the column valve)
• Post column pressure monitor (outlet of the column valve)
Monitors
27 /GE 28-9577-83 /
Version2.0
UV monitor
Enables continuous measurement of the UV absorbance
Function• Monitor the UV absorbance of
buffer and eluted proteins• Flip-mode (3 wavelength)
Details• 190 - 700 nm• Flow cell
– 2 mm path length (default)– 10 mm path length
• True path length in each flow cell
Monitors
28 /GE 28-9577-83 /
Version2.0
ÄKTA™ avantSetting the future industrial standard
Inspired speed
卓越的安全性
Inspired scalability
29 /GE 28-9577-83 /
Version2.0
Security
可靠的数据是深入掌握工艺流程的关键
Built-in fraction collector for true sample integrity
Reliable column protection with differential pressure
measurement
UniTag scanning for complete traceability
• Contaminant-protected and temperature controlled
• Capacity increased by 30% versus Frac-950
• Tube rack types are automatically detected
• Two pressure sensors protect the column and the media from overpressure
• Differential pressure measurement over columns
• Track column and run data easily with UniTag scanning
• Individual columns identified using a matrix code, key column performance parameters automatically saved in the column’s history
Working faster while being compliant
• UNICORN™ 6 is FDA 21 CFR Part 11 compliant and features a system audit trail, electronic signatures, and electronic records
30 /GE 28-9577-83 /
Version2.0
内置型组分收集器确保样品安全
ÄKTA™ avant has a built-in, temperature controlled fraction collector with automatic detection. Allows for hassle-free overnight running, outstanding reproducibility and sample security!
• 内置型组分收集器--无灰
• 相对封闭的收集腔--控温
• 收集管放于方便拿取的支架上--不会错
• 有7种不同的支架--灵活
• 可以自动识别支架类型--无误
• 防液滴滴出的功能--节约
• 比传统的 Frac-950增加30%的收集能力--高通量
Fraction collector
31 /GE 28-9577-83 /
Version2.0
Column handling 确保层析柱的操作安全
减少因层析柱性能降低、过期、误用等导致的失败运行,并对层析柱的使用进行追踪
• 确认: 层析柱的类型和参数
• 追踪: 层析柱的使用次数和使用情况
• 安全: 可以设置CIP和柱效测
定的频率,确保柱子的良好性能
Easy identification, automatic traceability
and outstanding security of operation!
UniTags
32 /GE 28-9577-83 /
Version2.0
文档保护,使用痕迹,电子签名确保数据安全
UNICORN™ 6 is suitable for use in a regulated environment in a manner complying with 21 CFR Part 11.
• System audit trial, electronic signatures, and electronic records.• Individual user access permissions• Lock the system according to a defined time schedule • All maintained records are stored in a single, unalterable database• Validation support documentation includes:
Detailed description of the development model used for UNICORN 21 CFR Part 11 system assessment Audit report and 21 CFR Part 11 conclusion on functionality by an external and
independent expert
See film clip on the following slide!
33 /GE 28-9577-83 /
Version2.0
柱前压力和压力差 (∆P)
For increased operational safety, the pressure difference over the packed media bed (∆P) is continuously measured.
• Pressure sensors integrated into the column valve protect the column and media from overpressure
• One pressure sensor measures the pressure before the column to protect the column hardware.
• The other sensor measures the pressure after the column and calculates the pressure difference over the packed media bed (∆P).
• Flow regulated mode automatically decreases flow rate when pressure exceeds the preset limit.
• Pressure sensors are also connected to the system and sample pumps.
See film clip on the following slide!
34 /GE 28-9577-83 /
Version2.0
ÄKTA™ avantSetting the future industrial standard
Inspired speed
Inspired security
可放大性
35 /GE 28-9577-83 /
Version2.0
卓越的可放大性
ÄKTA™ avant能确保工艺可以顺利地从研发阶段过渡到生产阶段
UNICORN™ 6软件操作平台 设计适合用新一代的BioProcess™ media
配合HiScreen™ 预装柱开发工艺,轻而易举的放大
• Built-in application knowledge for planning, executing, and analyzing chromatography at all scales
• Improved usability and visual design
• New functionality for column handling and statistics
• Superior management of methods, results, data, protocols, etc.
• Flow rate and pressure specifications that support BioProcess media such as MabSelect™ and Capto™,
• ÄKTA avant, together with BioProcess media, reduces process time with high volume throughput and transparent scale-up
• Designed for method optimization with the added flexibility of being able to translate to large scale production
• Prepacked with a range of modern BioProcess media
36 /GE 28-9577-83 /
Version2.0
使用高流速填料增加产能Media such as MabSelect™ and Capto™ are specifically designed to meet the demands of today’s industrial biomanufacturing. ÄKTA™ avant has flow rate and pressure specifications that support these modern, high flow BioProcess™ media, allowing for smooth scale-up
• Direct scalability to process scale, using the same media
• Increased productivity with high dynamic binding capacity at high flow
• Increased yield with rapid mass transfer
• Reduced process time with high volume throughput
• Cost-effective processing with smaller unit operations
Full support of modern media…Screening, column id 5-16 mm: 20-25 ml/minScale-up, columns id 16-50 mm: 120-130 ml/min
37 /GE 28-9577-83 /
Version2.0
Process economy gains with modern resins …
10.911.8
19.2
37.4
0
5
10
15
20
25
30
35
40
Classic process Protein A update Protein A 3-stepmodel process
Protein A 2-stepmodel process
Dow
nstr
eam
cos
t co
ntri
buti
on [U
SD/g
]
0
20
40
60
80
100
120
DSP
pro
cess
tim
e [h
]
Working volume 10,000 LTiter 5 g/LYield 79%
(Sepharose™ Fast Flow resins)
(MabSelect SuReCapto™ S Capto Q)
(MabSelect SuRe™Sepharose Fast Flow)
(MabSelect SuReCapto adhere)
• Cost savings with modern resins up to 70%
• Process time down to 2 days from 5 days
• 2-step process: additional indirect gains from space and tankage requirements
Use of modern resinscombined effect of capacity, flow, and lifetime
38 /GE 28-9577-83 /
Version2.0
专业专注的HiScreen预装柱ÄKTA™ avant is optimized with HiScreen™ pre-packed columns. Using pre-packed formats,Save a day’s work and make scale up easy!
• Use pre-packed formats and save time on packing , preparation of gel, hardware and solutions, column testing and possible re-packing
• 19 different BioProcess™ media - well characterized and robust, all regulatory support available
• Valco fitting – direct connection to ÄKTA™ design
• 10 cm bed height - ideal for method optimization and parameter screening, easily connected in series
• Scalable to BioProcess columns packed with the same media using the same process fluid velocities
Reliable, reproducible and easy to use!
Columns
39 /GE 28-9577-83 /
Version2.0
ÄKTA™ avantSetting the future industrial standard
Inspired technology
40 /GE 28-9577-83 /
Version2.0
System Orientation
Interactive instrumentdisplay delivers realtimeinformation.
Buffer tray for convenient,large capacity storage.
One wet side with clear view of flow path for convenience and usability.Module-
Module-based design including pumps,valves, and monitors.
Snap-on columnconnection providesadditional convenience
High precision systempumps with integratedair sensors.
Swivel foot for easy accessto all parts of the instrument
Climate-controlled,built-in fractioncollector preventslocal heating ofsamples.
Automatic sample applicationeliminates manual re-plumbing.
41 /GE 28-9577-83 /
Version2.0
System OrientationWith it’s built in fraction collector, ÄKTA™ avant has a smaller foot print and greater practical accessibility than its predecessor ÄKTAexplorer™
ÄKTA avant
860 mm 1100 mm
ÄKTApilot™
ÄKTAexplorer & Frac-950
ÄKTA avant
ÄKTAexplorer and Frac-950
42 /GE 28-9577-83 /
Version2.0
Setting the new industrial standardFor the last decade, ÄKTAexplorer™ has been the industry standard of process development. The proud ÄKTA™ lineage now sky rockets into the next generation with ÄKTA avant. Now we set the new industry standard.
ÄKTA™ avant 25 ÄKTAexplorer™ 10 and 100
Flow rate 0.001-25 ml/min 0.001-10 or 0.01-100 ml/min
Maximum back pressure 200 bar, 20 MPa, 2900 psi50 bar, 5 MPa, 725 psi*
250 bar, 25 MPa, 3625 psi100 bar, 10 MPa, 1450 psi
Quaternary gradient √ -
Similar design of all pumps √ (system and sample pumps similar)
- (different sample and system pumps)
Inbuilt reversed flow √ √ (reversed flow valve required for ÄKTAexplorer)
BufferPro (truly automated buffer preparation with corresponding acid and base)
√ -
integrated frac. Sample protection, √ (inbuilt frac) - (separate frac. required)
Temp. conditioned frac. √ -
Automatic id. of optical path length (UV) √ -
DoE as part of UNICORN √ -
Phase editor √ -
Undo button in UNICORN √ -
Pre and post column pressure √ -
Delta pressure measurement √ -
Flow control to avoid overpressure √ -
Constant pressure flow √ -
Injection techniques (built in flow path) 3 1
pH calibration and storage (inbuilt) √ -
Restrictor selection automatic √ -
Rack validation in fraction collector √ -
Customized fraction collector configurations √ -
Columns labeling and recognition (UniTags) √ -
43 /GE 28-9577-83 /
Version2.0
Reliable column connectionsIn addition to columns and tubing, other ÄKTA™ avant accessories include holders and clamps for attaching columns, flasks, and tubing to the system
• Easy snap-on column connection and pre-packed screening columns (10 cm bed height)
• New column holder design that eliminates screwing when connecting columns.
Column connections
*
* To be released at a later date
44 /GE 28-9577-83 /
Version2.0
ÄKTA™ avant
• Shorten process time from from 5 to 2 days with modern resins
• New user interface in UNICORN, save 1 day’s work with easy and flexible method creation and reduce learning time to less than half!
• DoE significantly reduces your development costs
• Save 30 min per buffer and pH value with BufferPro
• Save 1 day’s work with system & sample pump set up to run independently & with no need for replumbing
• Save 1 day’s work with pre-packed columns
• Limited service down time with easy accessible wet side & modular set up
• Save time by running safely over night
• Save 30 min per week on pH calibration
ÄKTA avant reduces weeks to days…
… while giving you outstanding scalability and sample security
45 /GE /
Version2.0
GE Healthcare Ltd, a General Electric Company. GE, imagination at work and GE Monogram are trademarks of General Electric Company.
ÄKTA, ÄKTAavant, ÄKTAexplorer, ÄKTApilot, BioProcess, MabSelect, MabSelect SuRe, Capto, HiScreenand UNICORN are trademarks of GE Healthcare companies.
“MabSelect SuRe Ligand Restricted License” and “Cys-rProtein A Ligand Restricted License” are protected by the following patents and equivalent patents and patent applications in other countries: US 5,151,350, US 5,143,844, US 6,399,750, WO 03/00475and EP 1123389. A free, non-transferable limited license to use this product for internal analytical purposes only accompanies the purchase of the product from a GE Healthcare company and its licensed distributors. Any other use will require a separatelicense from a GE Healthcare company.
All goods and services are sold subject to the terms and conditions of sale of the company within GE Healthcare which supplies them. General Electric Company reserves the right, subject to anyregulatory and contractual approval, if required, to make changes in specifications and features shown herein, or discontinue the product described at any time without notice or obligation.
© 2009 General Electric Company – All rights reserved.
GE Healthcare Bio-Sciences AB, a General Electric Company.
GE Healthcare Bio-Sciences AB, Björkgatan 30, SE-751 84 Uppsala, Sweden.