Aspen simulation Gel filtration(GF) - CHERIC · 2009-12-17 · BSABSA&&NaClNaClPropertiesProperties...
Transcript of Aspen simulation Gel filtration(GF) - CHERIC · 2009-12-17 · BSABSA&&NaClNaClPropertiesProperties...
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Gel filtration(GF)Gel filtration(GF)Aspen simulation
Gel filtration(GF)Gel filtration(GF)
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ContentsContents
GFGFBSA&NaCl PropertiesColumnExample ConditionExample ConditionIP OptimizationA Si l ti V i blAspen Simulation Variables
Sample VolumeFlFlowScale up
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GF (Gel GF (Gel FilterationFilteration))
겔을 운반체(캐리어)로 사용하여 측정하고자 하는 고분자용액을 주입하면 분자량이 작은 분자는 겔이 이루고 있는 그액을 주입하면 분자량이 작은 분자는 겔이 이루 있는물에 잘 흡착되고, 분자량이 커질수록 잘 흡착되지 않는다.
유출시간의 차이에 의해서 고분자의 분자량 분포 측정이나유출시간의 차이에 의해서 고분자의 분자량 분포 측정이나, 분자량에 따른 분별에 사용된다.
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[그림 1] GF의 원리[그림. 1] GF의 원리
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BSABSA &&NaClNaCl PropertiesProperties
BSA (B i lb i ) N ClBSA (Bovine serum albumin)
소 혈청 알부민분자량 약 66 4 kD
NaCl
소금분자량 : 약 66.4 kDa종류 : 단백질 (대부분의 동물에 많이 존재)용도
분자량 : 58.44 Da용도
조미 염장 등의 일상 생활과용도- 세포 배양시에 세포의 영양분- 단백질 정량에서 검정곡선을 얻기위한 표준물
- 조미, 염장 등의 일상 생활과
공업 방면에서 사용
- 동물의 체내 삼투압의 유지위한 표준물
- 생화학적 실험에서 특정 항체를 검출 하고자 하는 단백질이 붙여주기 전에 nonspecific binding
동물의 체내 삼투압의 유지
는 단백질이 붙여주기 전에 nonspecific binding에 항체가 달라 붙는 비특이적 결합을 막아 주기위해 사용
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[그림. 2] BSA의 분자 구조와 3차원 리본 구조
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[그림. 3] NaCl의 화학 구조와 현미경으로 본 NaCl
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ColumnColumn
Column volume 5 ml
M di S h d ™ G 25 Medium Sephadex™ G-25 Superfine
Column dimensions 1.6 × 2.5 cm
Sample volume 0.25-1.5 ml
Max. flow rate (H2O at 25°C)
15 ml/minO at 25 C)
Recommended flow rate
1-10 ml/min
Max. pressure 3 bar (43 psi, 0.3 MPa)
HiTrap Desalting Column 5ml
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Experimental ConditionsExperimental Conditions
Column : HiTrap Desalting 5ml/Sample : Bovine serum albumin (BSA) 2mg/ml
0.5 M sodium chloride (NaCl)0.05 M sodium phosphatepH 7.0
Buffer : 0.05 M sodium phosphate0.15 M sodium chloridepH 7.0
Sample volume : 0.8 mlSample volume : 0.8 mlFlow : 5 ml/mim
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IPIP ChangeChange
1차 2차 3차(최종값)(최종값)
BSA IP1 1.0 0.5 0.1
NaCl IP1 2.0 3.0 2.5
BSA IP2 0.0 0.0 0.0
NaCl IP2 0.0 0.0 0.0
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Simulation VideoSimulation Videohttp://blog.naver.com/rapy1004/150075292689http://blog.naver.com/rapy1004/150075292689
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Simulation Video Simulation Video ‐‐ IP changeIP changegghttp://blog.naver.com/rapy1004/150075491308http://blog.naver.com/rapy1004/150075491308
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Aspen Simulation Variables Aspen Simulation Variables
Sample Volume0.8, 1.3, 1.7, 2.2 ml
Column Volume : 5 mll l/Flow : 5 ml/min
Flow1.7, 3.3, 6.7, 10.0, 16.7, 20.0 ml/min
Column Volume : 5 mlS l V l 0 8 lSample Volume : 0.8 ml
Scale upl l 3 l lHiTrap Desalting 1x5 ml, 3x5 ml, 5x5 ml
Sample : 28% x Vt (1.4, 4.3, 7.1 ml respectively)
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Sample VolumeSample Volume
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Simulation Video Simulation Video ‐‐ Sample VolumeSample VolumeSimulation Video Simulation Video ‐‐ Sample VolumeSample Volumehttp://blog.naver.com/rapy1004/150075491442http://blog.naver.com/rapy1004/150075491442
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Sample volume changeSample volume change
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FlowFlow
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Si l i idSi l i idSimulation Video Simulation Video ‐‐ FlowFlow
http://blog.naver.com/rapy1004/150075491634http://blog.naver.com/rapy1004/150075491634p // g / py 4/ 5 7549 34p // g / py 4/ 5 7549 34
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Flow rate changeFlow rate change
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Scale upScale up
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Simulation Simulation –– ScaleupScaleuppphttp://blog.naver.com/rapy1004/150075491739http://blog.naver.com/rapy1004/150075491739
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Scale upScale up
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ConclusionsConclusions
체류시간 및 용출시간과 관련된 변수는 IP, Column Volume FlowVolume, Flow
그래프 넓이 및 높이와 관련된 변수는 Sample VolumeVolume