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生物分子經常是經由溶劑梯度的批次層析來進行純化的,
SMB的主要缺點是它沒辦法把混合物分開成3個餾份, 也缺乏溶劑梯度的應用能力.
MCSGP 程序(多分離管逆流溶劑梯度純化) 把這兩種技術結合在一個程序中,
即逆流SMB原理及溶劑梯度批次技術.
目前最完整地發展及提供 MCSGP 技術及設備的是瑞士的 ChromaCon 公司.
http://www.chromacon.ch/ 的
Contichrom ® lab (10/100m/min) 系統.
Contichrom ® 系統的靈活程度, 允許用一個單一的設備部件及簡單的過序轉換,
開發所有下游的各種流程模式, 包括 :
批量(batch)
模擬移動床層析(SMB)
補集式模擬移動床層析(CaptureSMB)
多分離管列(multi-column)
多分離管逆流溶劑梯度純化(MCSGP)等.
Contichrom ® lab (10/100m/min) 系統的全球銷售及服務則由歐系最具規模的HPLC製造廠,
德國 Knauer 公司負責提供.
http://www.knauer.net/en/products/product-lines/contichrom-purification.html
ABDC 則是Knauer及ChromaCon 公司的主要經銷者之一,
ABDC-CXTH 則是 亞洲最大自動純化設備研發, 製造及服務團隊,
也是Knauer 公司的大陸及香港總代理.
http://www.chromnet.net/PreparativeChromatography.aspx
以下是
壹. Contichrom ® lab (10/100m/min) 系統簡介,
貳: 維基百科的 多分離管逆流溶劑梯度純化(中英對照).
壹. Contichrom ® lab
(10/100m/min) 系統簡介(中英對照)
1. How can flexibility and process economics be combined in one LC system?
靈活度與經濟的流程如何能結合在一個LC系統？
2. Contichrom ® lab equipment for superior LC purification
Contichrom ® 優越的液相層析純化實驗室設備
Which aspects are important for a purification system?
對於一個純化系統, 那些方面是重要的 ?
Introducing the flexible Contichrom ® lab (all-in-one) process capability.
導入靈活的Contichrom ®實驗室(全部合一)的流程能力.
Flexibility of Contichrom ® allows developing all downstream process modes
(batch, SMB, CaptureSMB, multi-column, MCSGP) with one single piece of equipment
and easy process conversion
Contichrom ® 的靈活程度允許用一個單一設備及簡單的流程轉換, 開發所有下游的各種流程模式
(批量, 模擬移動床層析, 補集式模擬移動床層析, 多分離管列, 多分離管逆流溶劑梯度純化).
Process economics
The MCSGP process increases both purity and yield by 50 % compared to the
corresponding batch process
• Contichrom ® allows rapid automated process-development
經濟的流程與對應的批處理過程相比較, MCSGP流程同時增加了50 ％純度和產率.
• Contichrom ®允許快速自動化的流程開發
Scalability
Contichrom ® allows retention of the developed product profile, yield and purity
specifications upon upscaling
可擴展性
在尺度向上提升時, Contichrom ®允許保留已開發產品的輪廓調件, 產量和純度規格等.
Contichrom ® lab is a preparative liquid chromatography system designed for
discovery, process development and scale-up, offering great flexibility in
process choices. With Contichrom ® lab, all process modes (batch, SMB,
CaptureSMB, MC- SGP, multi-column) can be performed with a sin- gle equipment
and control software.
Contichrom ®實驗室是一個製備液相層析系統, 設計用於探索發現, 處理過程的發展及放大,
在處理過程的選擇上提供很大的靈活度. 有了Contichrom ®實驗室, 所有各種的處理過程模式(批量,
模擬移動床層析, 補集式模擬移動床層析, 多分離管逆流溶劑梯度純化, 多分離管)
都可以用一個單一設備和控制軟體來進行.
The Conti-chrom ® control software has been developed to enable the conversion
of batch to multi-column processes using a method wizard and to allow the fast
development of scalable processes, omitting extensive screening steps.
Contichrom ® 控制軟體已經開發出來, 使其能夠使用方法精靈嚮導進行由批次到多分離管的轉換,
並允許可擴展的流程的快速發展, 省去廣闊範圍及深度的篩選步驟.
The Contichrom ® lab adds another dimension to process chromatography, as it
allows develop- ment of processes based on the MCSGP (Multi- column
Countercurrent Solvent Gradient Purifica - tion) principle which is much more
efficient than a batch process.
All standard media (Ion exchange, mixed-mode, HIC, RP, SEC, Affinity) can be
used as well as all standard organic solvents and aqueous buffers.
Contichrom ®實驗室增加了另一個層面層析處理, 因為其允許開發基於MCSGP(多分離管逆流溶劑梯度純化)原理的流程, 它比批次程序更有效率.
如同所有標準的有機溶劑和水性緩衝液等,
所有標準介質(離子交換, 混合模式, HIC, RP, 斥濾層析, 親和層析) 都可以使用.
Which processes can your purification system operate?
Flexibility in process development means that based on the purification
challenge the best process is chosen with respect to ease, development time and
costs.
您的純化系統可以運作哪些流程 ?
流程發展的靈活性指的是基於對於簡單的程度, 開發的時程和成本, 選擇最好的流程的淨化的挑戰.
For affinity capture with low dynamic binding capacity having a broad
breakthrough curve, CaptureSMB is superior to batch.
For simple ternary separations, batch processes are the best options if product
(red) and impurity (grey) are well separated.
However for difficult ternary separations of overlapping impurities, the MCSGP
process is superior.
對於具有低的動態聯結容量, 具有寬的穿透曲線的親和捕獲, 親和捕獲SMB優於批次層析.
對於三元分離, 批次層析是最好的選項, 如果產物(紅色) 與不純物(灰色)分開的很好
但不論如何, 對於困難的重疊不純物的三元分離, MCSGP 流程是較優的.
The isolation of product-related impurities for pre-clinical and toxicological
assessments, nor- mally a tedious task, is easy using the MCSGP process(far
left).
With Contichrom ® lab equipment all processes as shown above can be performed.
臨床前和毒理評估上的產物相關雜質的分離, 通常是件乏味繁索的工作, 用MCSGP流程是很容易的(最左邊).
用Contichrom ®實驗室設備如上所示的所有過程能被呈現出來.
Contichrom ® lab equipment allows development of a simple batch process first
and then automatic switching to an MCSGP process, providing superior process
economics.
Contichrom ®實驗室設備允許先發展一個簡單的批流程, 然後自動切換到MCSGP過程中, 提供卓越的流程的經濟性.
How can affinity capture be faster and more cost effective?
Using CaptureSMB ® , a twin column cyclic process, with Contichrom ® equipment
instead of a single column process for Protein A capture steps has significant
advantages:
親和捕獲如何更快和更具成本效益？
使用CaptureSMB ® , 一個雙生分離管循環程序, 與Contichrom ®裝備 - 而不是單分離管程序, 於蛋白A捕獲步驟有顯著的優點:
• Higher loading velocities shortening the transit time for the capture step by
50 %
• Due to faster processing, preservation of protein integrity
• Optimal use of Protein A resin capacity leading to a resin cost saving of 50 %
• Annual Protein-A resin cost savings in production scale of several Mio USD
• Easy and robust twin-column equipment design and software wizard
•更高的裝載速度, 縮短捕獲步驟的過渡時間50 ％.
•由於更快的流程, 保存蛋白質的完整
•蛋白質A樹脂容量的最佳利化用, 達到樹脂的成本節省50 ％
•在生產規模下, 節省幾百萬美元的蛋白A樹脂年成本
•簡單和強大的雙分離管設備的設計和軟體引導精靈
Application Areas
• Fast capture of monoclonal antibodies
• Capture of proteins to customized ligands (chemical and bilogical)
• Capture of therapeutic compounds from fractionated blood plasma
• Capture of API from natural extracts
應用領域
•單株抗體的快速捕獲
•特製配體(化學和生物的)的蛋白質捕捉
•分餾後的血漿的治療用化合物捕獲
•天然提取物中的API(活性藥物成份)捕獲
How can purity and yield be optimized simultaneously?
純度和回收率如何同時優化？
The MCSGP process operates with twin columns instead of one column, whereby the
impure side fractions containing product are recycled inter- nally, extracting
all product and thereby increa- sing both yield and purity.
The MCSGP process mode is typically operated with the same transit time as a
batch chromatography step.
此 MCSGP 流程以雙分離管操作, 而不是一支分離管, 含有不純物側的餾分的產物在內部被回流, 因而能提取出所有的產物, 增加產量和純度.
MCSGP程序模式通常以操作跟批次層析步驟相同的過渡時間操作.
A batch derived from a MCSGP process has the same «batch definition» as one from
a normal single column batch process.
It is a specific quantity of material or API processed in one process or series
of processes so that it could be expected to be homogenous.
從MCSGP程序中導出的一批次, 與從正常的單分離管批流程有同樣的«批次的定義».
它是在一個進程或一系列的流程裏, 一個被預期可以得到均質的特定被處理的材料或原料藥的量.
Conventional batch chromatography
傳統的批處理層析法
impure product to waste
棄置的不純產品
pure product
純品
ChromaCon's internal recycling chromatography (MCSGP)
ChromaCon的內循環層析法 (MCSGP)
Reprocess impure product
重新處理不純產品
Cut narrow = obtain purer roduct more and purer product
切窄=獲得更純淨的產品更多和更純的產品
Application Areas
• Discovery: isolation of leads
• Process development: purification of proteins, synthetic peptides
• Purification of oligonucleotides
• Isolation of product-related impurities
• Purification of conjugates and PEGylated proteins
• Purification of small molecules and natural products
應用領域
•發掘：分出前導物
•流程開發：蛋白質的純化, 合成肽
•寡核苷酸的純化
•與產品相關的雜質的分離
•共軛物和聚乙二醇化蛋白的純化
•小分子和天然物的純化
How can impurities be isolated more efficiently?
Applications in Discovery and in the isolation of product-related impurities:
The Problem:
• Discovery:
In shotgun proteomics (bottom-up), due to the complexity of the samples,
extensive front-end separation is essential to alleviate the problem of peptide
undersampling, to maximize protein identifications and to tackle the vast
dynamic range.
怎樣雜質才能被更有效地分離？
在發掘及在與產品相關的雜質的分離的應用：
問題：
•發掘: 在鳥槍法蛋白質體學(自下而上)上, 由於樣品的複雜性, 廣泛的前端分離是必不可少的,
以能減輕肽取樣不足量的問題, , 以最大化蛋白質鑑定, 並承擔廣大動態範圍.
• Related impurities:
Isolation of product-related impurities is normally a tedious task requiring
several hundred injections to isolate sufficient material for pre-clinical and
toxicological studies.
The Solution:
• The MCSGP process allows selective enrichment of a defined region of the
chromatogram (red)
• The enriched fraction can be isolated for functional characterization or
analyzed directly online using mass spectrometry analysis Selectively enriched
fractions
•相關雜質：
產品相關雜質的隔離, 通常是一個需要幾次百注射以分離出足夠用於臨床前和毒理學研究材料的單調乏味的任務.
解決方案：
•MCSGP流程允許在一個層析圖的定義區域裏 (紅色)選擇性地富集.
•富集的餾分可被分離, 用於 功能鑑定或直接地在線上進行質譜分析選擇性富集餾分
Does your system provide automated process development?
您的系統提供自動化流程的發展嗎？
Contichrom ® lab equipment and software have been designed to be very
user-friendly.
The control software helps the user to easily navigate and manage process setup,
execution, and reporting.
Fast and automated process development
• Wizards with graphical user interface for easy method programming
• Automated conversion from batch to MCSGP process
• Extensive library of pre-defined methods for all standard operations
Contichrom ®實驗室的設備和軟體已經被設計得非常使用者友好. 控制軟體可以幫助用戶輕鬆地瀏覽和流程設置,
執行和報告.
快速和自動化流程開發
•精靈嚮導具有簡單的方法編程的圖形使用者界面
•從批量到MCSGP流程的自動轉換
•對所有標準操作預先定義好方法的廣泛資料庫.
Easy to operate
• Intuitive software for operation of batch and MCSGP
• Active flow path highlighted in flowsheet
• Pause/continue functionality, even for continuous chromatographic operations
Integrated evaluation and reporting
• Detailed evaluation capabilities with standardized PDF reports
• Data export functions
• Full audit trail and change control
• User management hierarchy provides high operational and data security
• FDA 21 CFR Part 11 compliant
Batch wizard
Batch to MCSGP wizard Main window Method editor Evaluation & report center
System online monitoring Flowsheet and active flowpath
綜合評價和報告
•標準PDF報告的詳細評估能力
•數據導出功能 Full data security and traceability
完整的數據安全性和可追溯性
•完整的審計記錄和變更控制
•用戶階層管理, 提供高度操作和數據保全
•符合FDA 21 CFR Part 11的標準
批次精靈嚮導
批次轉 MCSGP 精靈嚮導主視窗方法編輯器評估與報告中心系統在線監控流程圖和動作路徑
易於操作
•批次和MCSGP流程操作的直觀軟體
•動作中的流動路徑凸顯在流程表中
•暫停/繼續功能, 即使是連續的層析操作
How can you switch to a superior process?
您能如何切換到卓越的流程？
The Contichrom ® wizard allows switching a batch process to a superior MCSGP
process in just three steps:
Step 1:
Firstly, a non-optimized batch process is developed, the product peak is
identified and the batch chromatogram is stored in the database. For conversion
of a batch to an MCSGP process, the respective batch chromatogram is retrieved.
Step 2:
Using the Contichrom ® software wizard, the borders of the product peak (red)
are defined by simply dragging bars to define the boundaries. The narrower the
boundaries are defined, the purer the isolated product. Then the boundaries of
the recycling fractions (blue) are set by dragging another set of bars and the
start and end points are defined.
Step 3: Pressing the software wizard button will automatically convert batch
process into an MCSGP process.
Contichrom ®精靈嚮導允許只需三個步驟, 即可切換批次流程到卓越的MCSGP：
步驟1：首先, 一個非優化的批次流程過程被開發, 其產品峰被識別和批次層析圖被存儲在數據庫中. 轉換一個批次流程到一個MCSGP流程時, 批次層析圖被取用.
第2步：使用Contichrom ®軟體精靈嚮導, 產物峰(紅色)的邊界被簡單地以拖曳圖控拉桿來定義其界限. 所定義的邊界愈窄, 所分離的產品愈純. 然後,
要回流的餾份(藍色)的界限, 由拖另一組圖控拉桿來設定, 並且其起始點和結束點被定義了.
第3步：按軟體精靈嚮導按鍵, 會自動轉換批次流程到一個MCSGP 流程.
Step 1:
retrieve chosen chromatogramm of batch run from database
Step 2: interactive definition of product range (red) and recycling fractions
(blue): pull bars to define boundaries (dotted lines)
Step 3: press button to convert batch to MCSGP process
步驟1：取用從資料庫選擇的批次流程層析圖
第2步：產品範圍(紅色)和回流部分(藍色)的互動式地定義：拖曳圖控拉桿以定義界限(虛線)
第3步：按下按鈕以轉換批次到MCSGP 流程.
How do you combine multifunctionality with a small footprint?
您如何結合多重功能與小的足跡？
Application fields:
• Process development: facilitated through flexibility in choosing the right
process for a given process challenge
• Isolation of product-related impurities:
the tedious task of isolating product-related impurities for pre- clinical /
toxicological characterization is considerably facilitated using Contichrom ®
with MCSGP mode
•Preparative product purification: The Contichrom ® lab is available in two
versions with two different pumps:
•Contichrom ® lab-10: 0.01–10 mL/min for up to 20 g product /day
•Contichrom ® prep-100: 1–100 mL/min for up to 200 g product /day 應用領域：
•流程開發：對於給定處理程序的挑戰, 經由選擇合適的流程的靈活度而被促進
•產品相關的雜質的分離：在臨床/毒理學定性方面, 分離產品相關雜質的繁瑣任務, 使用Contichrom®與MCSGP模式是大大地被促進.
•製備產物純化：Contichrom ®實驗室有兩個不同的泵的版本可供選擇：
• Contichrom ®實驗室- 10 ： 0.01〜10毫升/分鐘, 最多20克產品/天
• Contichrom ®準備- 100 ： 1-100毫升/分鐘, 高達200克的產品/天
High precision
sample pump
UV detectors with low hold-up volume
Precise high-pressure gradient pumps
Reliable multi-position valves
The Contichrom ® purification system requires less space than most preparative
HPLC systems.
高精度
採樣泵
具有低滯留量的紫外檢測器
精確的高壓梯度泵
可靠的多位閥
Contichrom ®淨化系統需要比大多數製備型高效液相層析系統少的空間.
Multi-functionality with a small footprint
多重功能與小的足跡
• Contichrom ® is a preparative liquid chromatography system intended for
process screening and development.
• Instead of time-consuming Design of Experiments (DoE) for establishing
optimized starting conditions, Contichrom ® offers a software wizard tool to
easily convert a basic batch process to a superior MCSGP process, thus reducing
development time for robust and economic processes considerably.
• Contichrom ®是一種製備型液相層析系統用於流程的篩選和開發.
• 相反於為了建立優化的初始條件的耗時的試驗設計, Contichrom ® 提供了一個軟體精靈嚮導工具以輕鬆地把基本的批次流程轉換到優異的MCSGP流程,
從而相當程度地減少了強勁及經濟流程的開發時間.
• The Contichrom ® operating software provides pre-defined methods for standard
batch operations and for an intuitive user-friendly operation and data security,
being compliant to 21 CFR part 11 requirements. • The Contichrom ® lab has a
modular design where parts can be easily accessed and replaced.
It has an interactive instrument display and many functionalities, such as a
pause / continue functionality. A large buffer storage area on top of the
instrument provides sufficient space for continuous process modes.
•Contichrom ®操作軟體提供了用於標準的批次操作和一個直覺的使用者友好的操作和數據的安全性的預設方法, 符合21 CFR Part 11的要求.
•Contichrom ®實驗室 採用模組化設計, 其部件可以很容易地存取和替換. 它有一個互動式的儀表顯示及許多功能, 例如一個暫停/繼續功能.
在儀器的頂部有一個大的緩衝液儲存區提供用於連續流程模式足夠的空間.
• The system and sample pumps are based on high quality KNAUER BlueShadow ® pump
technology, ensuring reproducible flow rates at both low and high pressures.
• The system is suitable for low, medium and high pressure and is solvent
resistant and biocompatible, offering functionality for small molecule and
macromolecule preparative purifications.
•系統和樣品泵是基於高品質的 KNAUER BlueShadow ®泵技術, 確保在低和高的壓力校的可再現流率.
•系統適用於低, 中, 高壓力, 而且是溶劑耐受性和生物相容性, 提供了對於小分子和大分子製備純化的功能.
• The Contichrom ® system is scalable to pilot and process scale. At up-scaling,
the product profile is retained.
• The Contichrom ® system is particularly suitable for PAT in line with QbD.
•Contichrom ®系統是可放大到前導和製程的規模. 在規模放大時, 產品輪廓是被保留下來的.
•Contichrom ®系統特別適合於用QbD的在線PAT .
Contichrom ® Technical specifications
Main system components:
• System configuration: 2-column setup
• 1 x binary high pressure gradient pump
• 2 x isocratic pumps, 2 x UV detectors
• 6 x 6-way 7-port valves, 2 x 2-way 6-port valves
• 1 x degasser, 1x pH / conductivity sensor
• 1 x computer / monitor / keyboard / mouse
• 1 x Contichrom ® operating software
• 1 x ethernet switch
技術規格主要的系統組件：
•系統配置：2-分離管的設置
•1個二元高壓梯度泵
•2個等度泵之, 2個紫外偵檢器
•6個 6向7通閥, 2個2向6通閥
•1個脫氣機, 1個pH值或電導率偵檢器
•1×電腦/顯示器/鍵盤/鼠標
•1個Contichrom ®操作軟體
•1個以太網連接器
Maximum system pressure: 100 bar Pump description:
• double piston pump
• biocompatible, titanium inlays
• active piston seal wash
• flow rate: • Contichrom ® lab-10: 0.01–10 mL/min
• Contichrom ® prep-100: 1–100 mL/min
• maximum delivery pressure: 100 bar
• gradient capability
最大系統壓力： 100 bar 泵說明：
•雙活塞泵
•生物相容性, 鈦嵌體
•主動活塞密封墊沖洗
•流量：
• Contichrom ®實驗室-10 ： 0.01〜10毫升/分鐘
• Contichrom ®準備- 100 ： 1-100毫升/分鐘
•最大輸出壓力： 100巴
•梯度功能
UV detector description:
• spectrophotometer for routine HPLC and other flow-through detection tasks
• wavelength range 190 –750 nm
• light source: deuterium lamp Degasser description (lab-10):
• 4-channel degasser module • degassing by gas permeation through Teflon ® AF
amorphous fluoropolymer membrane
紫外檢測器的描述：
•分光光度計用於常規HPLC和其他 流通方式的檢測任務
•波長範圍190 -750奈米
•光源：氘燈 脫氣描述(實驗室-10)：
• 4通道脫氣機模組
•脫氣 以氣體滲透通過Teflon ®AF非晶含氟聚合物膜
Valve description:
• electrical valve drive
• LED display
• PEEK body, TEFZEL seal
pH / conductivity sensor description:
• Conductivity 0–500 mS/cm
• pH measurements 0–14
• temperature sensor
閥描述：
•電氣閥門驅動器
• LED顯示器
• PEEK閥體, TEFZEL 密封
pH值/電導度率偵檢器描述：
•電導度0-500毫西門子/厘米
•測量pH值 0-14
•溫度偵檢器
Materials:
• all biocompatible
• tubing: PEEK
• Contichrom ® lab-10: 0.5 mm i.d.
• Contichrom ® prep-100: 1.0 mm i.d.
Fraction collector (optional):
• Fraction collector FOXY R1 ISCO with integrated diverter valve
• Fraction capacity from 36–144 tubes, 0.2 mL–50 mL per fraction
• biocompatible and can be used in cold rooms
材料：
•全部為生物相容性
•管路： PEEK
• Contichrom ®實驗室-10：0.5毫米內徑
• Contichrom ®準備- 100：1.0毫米內徑
餾分收集器(可選) ：
•餾分收集器 FOXY R1 ISCO 與分流閥整合
•餾分容量從36-144管, 每餾分0.2毫升-50毫升
•生物相容性, 可以在冷藏室中使用
Purified product output capacity:
• Contichrom ® lab-10: up to 20 g/day
• Contichrom ® prep-100: up to 200 g/day
Operation:
• room temperature
• cold room (4 °C) compatible
純化產物輸出容量：
• Contichrom ®實驗室- 10 ：高達20克/天 •
Contichrom ®製備- 100 ：高達200克/天
操作：
•室溫
•冷藏室( 4℃)兼容
Contichrom ® Operating software features
Software description:
• Contichrom ® operating software (in English) including manual.
• Integrated, intuitive software for operation and evaluation of batch and
continuous chromatography.
• Wizard allows easy programming of methods including MCSGP.
Contichrom ®操作軟體功能軟體描述：
• Contichrom ®操作軟體(英文)包含手冊.
•整合的, 用於操作和批次和連續層析的評估的直覺軟體.
•精靈嚮導允許輕鬆地進行方法的編程, 包括MCSGP.
• Run log stores all information for each experiment.
• Easy-to-use graphical user interface for method programming. Advanced users
can access program detailed settings.
• Unique pause / continue functionality, even for continuous chromatographic
operations.
•運行日誌為每個實驗存儲所有信息.
•用於方法編程上的易於使用的圖形使用界面. 進階使用者可以存取編程上的詳細設定.
•獨特的暫停/繼續功能, 即使是用於連續層析操作.
• User management gives high operational and data security.
• Provides all tools for FDA 21 CFR Part 11 compliance. Pre-defined user groups
• Administrators
• R&D-, production-users Rights management
• Each group has defined rights as set by the administrator
• User accounts are password protected Logging
• All user/hardware activities are logged with time stamp and user name
Electronic signature
• A checksum of the log & measurement file is computed and saved together with the files in order to
create an electronic signature
•用戶管理提供了很高的操作和數據的安全性.
•提供美國 FDA 21 CFR Part 11 規範的所有工具. 預先定義的用戶組
•管理員 •研發, 生產用戶權限管理 •每個組有由管理員設定的定義權限
•用戶帳戶密碼保護日誌
•所有用戶/硬體活動都有帶時間標記和用戶名的記錄電子簽名
•日誌和量測檔案的編碼加總校驗被計算, 並與這些檔案一起保存, 以產生電子簽名.
Operating software:
main window: status & flowsheet tab Main-Flowsheet Tab
• active connections are highlighted
• display of status / measurement of
• valve position
• pump parameters
• UV signals / settings
• conductivity
• pH / temperature
• tank levels
• run time informaion
• method in operation
• duration of time step
操作軟體：主視窗：狀態和流程標簽頁主流程表標簽頁
•連接動態亮度突顯
•狀態/測量的顯示
•閥門位置
•泵參數
• UV信號/設定
•電導度
• pH值/溫度
•儲存桶液位
•運行時間資訊
•操作的方法
•時間步驟的持續時間
When will you start Please contact us ...... for an offer, technical information
and support, after-sales service, shipping information:
您什麼時候開始請與我們聯繫......一個報價, 技術信息和支持, 售後服務, 發貨信息：
Wissenschaftliche Gerätebau Dr. Ing. Herbert Knauer GmbH Hegauer Weg 38 14163
Berlin Germany Phone: +49 30 80 97 27- 0 Fax: +49 30 8 01 50 10 www.knauer.net
contichrom@knauer.net ChromaCon and Contichrom are registered trademarks of
ChromaCon AG | All goods and services are sold subject to terms and conditions
of sale of KNAUER and ChromaCon. | Copies of these are available upon request. |
© 2013 ChromaCon AG – all rights reserved.
由於我們的協力原廠 CXTH也是 Knauer 及ChromaCon 大陸及香港總代理, 對於您未來的耗材及技術支援,
可以多一個很好的來源與保障的. 後續您要 Knauer ContiChrom 進行任何尺度的 Scale-UP
是更直接有效率,且成本低很多的.
http://www.chromnet.net/PreparativeChromatography.aspx
貳: 維基百科的 多分離管逆流溶劑梯度純化.(中英對照)
Multicolumn countercurrent solvent gradient
purification
(多分離管逆流溶劑梯度純化)
http://en.wikipedia.org/wiki/Multicolumn_countercurrent_solvent_gradient_purification
Multicolumn Countercurrent Solvent Gradient Purification
(MCSGP)
is a form of chromatography that is used to separate or purify
biomolecules
from complex mixtures. It was developed at the Swiss Federal
Institute of
Technology Zurich by Aumann and Morbidelli.[1]
The process consists of two to six chromatographic
columns
which are connected to one another in such a way
that as the mixture
moves through the columns
the compound is purified into several fractions.
多分離管逆流溶劑梯度純化(MCSGP)是一種用於從複雜的混合物中分離
及純化生物分子的層析的型態.
它是在瑞士蘇黎世聯邦科技學院的
Aumann and Morbidelli所發展出來的.
這種程序由2-6支分離管層析分離管所組成, 這些分離管連接的方式是使混合物通過這些分離管,
而化合物則是被純化到幾個部份.
Contents(內容):
Overview(綜覽)
The MCSGP process consists of several chromatographic columns which are
switched in position opposite to the flow direction. Most of the columns are
equipped with a gradient pump to adjust the modifier concentration at the column
inlet. Some columns are connected directly, so that non pure product streams are
internally recycled.
Other columns are short circuited, so that they operate in pure batch mode.
MCSGP程序由數支分離管層析分離管所組成,過閥門的切換, 以與流體流動的相反方向來變換這些分離管串接位置.
大部份的分離管都配備有梯度PUMP以調節分離管入口的修飾劑的濃度. 某些分離管是直接連接的, 使不純的產物串流可以在內部循環.
其他分離管是走捷徑連接的,使其能以單純的批次模式來操作.
The system is split into several sections, from which every section performs a
tasks analog to the tasks of a batch purification. These tasks are loading the
feed, running the gradient elution, recycling of weakly adsorbing site
fractions, fractionation of the purified product, recycling of strongly
adsorbing site fractions, cleaning the column from strongly adsorbing
impurities, cleaning in place and re-equilibration of the column to start the
next purification run.
All of the tasks mentioned here are carried out at the same time in one unit.
Recycling of non pure site fractions is performed in countercurrent movement.
The fully continuous process consists of at least 6 columns. Semi-continuous
operation with only 3 columns shows the same performance and it can even have a
higher productivity.
此系統可分開為幾個區段, 每一區段表現出類同於批次純化的各項任務. 這些任務有進料載入, 梯度的進行,
弱吸附節點的餾份的回流,純化產物的分餾,強吸附節點的餾份的回流分離管中強吸附雜質的洗清, 原位置做分離管洗清及再平衡以進行下一回的純化程序.
所有提到的這些任務是在一個單元裏同時進行. 不純節點餾份的回流是以逆流方向進行.
完整的程序至少由6支分離管組成. 由3支分離管組成的半連續的操作表現出相同的性能,甚至有更高些的產出.
Comparison with other purification methods
(與其他純化方法的比較)
Biomolecules are often purified via solvent gradient batch chromatography. Here
smooth linear solvent gradients are applied to carefully handle the separation
between the desired component and hundreds of impurities.
The desired product is usually intermediate between weakly and strongly
absorbing impurities. A center cut is required to get the desired pure product.
Often the preparative resins have a low efficiency due to strong axial
dispersion and slow mass transfer. Then a purification in one chromatographic
step is not possible. Countercurrent movement as known from the SMB process
would be required.
生物分子經常是經由溶劑梯度的批次層析來進行純化的. 在這樣的方式下,使用平順的溶劑梯度,小心的處理在所要的成份與數佰種雜質之間的分離.
所要的產物通常是介於弱的及強的吸附不純物的中間. 一個中心切割是需要用以得出所要的產物的.
通常製被型的樹脂由於有軸向強分散性,以及低的質量轉移, 所以, 要在一個層析步驟中做純話是不可能的. 因而,
SMB的逆流移動程序就成為所需要的方法.
For large scale productions and for very valuable molecules countercurrent solid
movement need to be applied to increase the separation efficiency, the yield and
the productivity of the purification. The MCSGP process combines both techniques
in one process, the countercurrent SMB principle and the solvent gradient batch
technique.
對於非常有價值的分子的大規模生產, 需採用逆流的固態移動, 以增加純化的分離效率,產率及產量. MCSGP
程序把這兩種技術結合在一個程序中, 即逆流SMB原理及溶劑梯度批次技術.
Discontinuous mode consists of equilibration, loading, washing, purification and
regeneration steps. The discontinuous mode of operation allows exploiting the
advantage of solvent gradients, but it implies high solvent consumptions and low
productivities with respect to continuous countercurrent processes.
An established process of this kind is the simulated moving bed technique (SMB)
that requires the solvent-consuming steps of equilibration, washing,
regeneration only once per operation and has a better resin utilization.
However, major drawbacks of SMB are the inability of separating a mixture into
three fractions and the lack of solvent gradient applicability.
非連續模式是由平衡, 載入, 清洗, 純化及再生等步驟所組成. 非連續操作模式允許能發揮溶劑梯度的優點, 但相對於連續逆流程序來說,指的就是高溶劑消耗及低產量.
一種已經建立的這種程序是以模擬移動床(SMB)方式建立的, 它在每一回操作只需要做一次的平衡, 清洗, 再生等溶劑消耗步驟.
但不論如何, SMB的主要缺點是它沒辦法把混合物分開成3個餾份, 也缺乏溶劑梯度的應用能力.
In the case of antibodies, the state-of-the-art technique is based on batch
affinity chromatography (with Protein A or Protein G as ligands) which is able
to selectively bind antibody molecules. In general, affinity techniques have the
advantage of purifying biomolecules with high yields and purities but the
disadvantages are in general the high stationary phase cost, ligand leaching and
reduced cleanability.
在抗體方面, 最先進的技術是基於批次的親和層析以 Protein A or Protein G為配體), 它能購選擇性地連結抗體分子.
一般來說,親和層析在純化生物分子方面,具有高產率及純度的優點, 但一般來說,也有高固定相成本, 配體淋溶,以及逐漸降低的可清洗能力.
The MCSGP process can result in purities and yields comparable to those of
purification using Protein A. The second application example for the MCSGP
prototype is the separation of three MAb variants using a preparative weak
cation-exchange resin. Although the intermediately eluting MAb variant can only
be obtained with 80% purity at recoveries close to zero in a batch
chromatographic process, the MCSGP process can provide 90% purity at 93% yield.
A numerical comparison of the MCSGP process with the batch chromatographic
process, and a batch chromatographic process including ideal recycling, has been
performed using an industrial polypeptide purification as the model system. It
shows that the MCSGP process can increase the productivity by a factor of 10 and
reduce the solvent requirement by 90%.[2]
用Protein A, MCSGP程序能達到與這些純化可相比較的純度及產率.
第二個MCSGP雛型的應用力例子是用一種製備用的弱陽離子交換樹脂來分離3個MAb的變體.
雖然在一個批次層析程序裏,直接沖滌出來的MAb變體整只能有80%的產率, 回收率接進於零, MCSGP程序能在93%產率下,能提供90%的純度.
一個MCSGP程序與批次層析程,以及與具有理想回流的批次層析程序的數據比較,已經以工業的多肽純化模型系統呈現出來.
它顯示MCSGP程序能增加10倍的產能及減少90%的溶劑需求.
The main advantages with respect to solvent gradient batch chromatography are
high yields also for difficult separations, less solvent consumption, higher
productivity, usage of countercurrent solid movement, which increases the
separation efficiency. The process is continuous. Once a steady state is
reached, it delivers continuously purified product in constant quality and
quantity. Automatic cleaning in place is integrated. A pure empirical design of
the operating conditions from a single solvent gradient batch chromatogram is
possible.
相對與於溶劑梯度的批次層析程序, 主要的優點有: 對於難分離的情況也有高的產率, 少的溶劑消耗, 較高的產量, 以及使用能增加分離效率的逆流固相移動等.
Applications(應用)
All chromatographic purifications and separations which are executed via solvent
gradient batch chromatography can be performed using MCSGP.
Typical examples are reversed phase purification of peptides, hydrophobic
interaction chromatography or for example ion exchange chromatography of
proteins or antibodies.
The process can effectively enrich components, which have been fed in only small
amounts. Continuous capturing of antibodies without affinity chromatography can
be realized with the MCSGP-process.[3]
所有經由進行溶劑梯度的批次層析的純化與分離都能用MCSGP方法呈現.
典型的例子有逆相的肽的純化, 蛋白質或抗體的疏水性交互作用層析或離子交換層析.
此程序能有效地富集這些進料當中只有少量的各個成份. 可以用MCSGP程序來達成不用親合層析法的抗體連續捕集.
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