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

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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

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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

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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.

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ÄKTA™ avant and UNICORN™ 6.0

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ÄKTA™ avant设置未来分离纯化的新标准

快速

Inspired security

Inspired scalability

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来自“速度”的压力

验证猜想

科研进度

产品上市

解放劳动

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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

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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!

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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

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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

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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!

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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

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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!

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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!

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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

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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

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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

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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

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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

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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

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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

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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

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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

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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

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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

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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

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ÄKTA™ avantSetting the future industrial standard

Inspired speed

卓越的安全性

Inspired scalability

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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

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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

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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

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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!

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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!

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ÄKTA™ avantSetting the future industrial standard

Inspired speed

Inspired security

可放大性

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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

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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

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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

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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

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ÄKTA™ avantSetting the future industrial standard

Inspired technology

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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.

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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

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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) √ -

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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

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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

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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.