Bio-rad Foresight™ Chromatography Columns, Prepacked Manuel d'utilisateur

Foresight™ Chromatography MediaFilter PlatesInstruction ManualCatalog Numbers732-4701 732-4712732-4703 732-4714732-4705

Technical Support: 1-800-424-6723 • www.bio-rad.com 5For optimal results a phase ratio in the range of 20–40 is recommended.9 To minimize the erro

6 Technical Support: 1-800-424-6723 • www.bio-rad.comFigure 1A depicts six time points. The adsorbed concentration and the liquid phase concentratio

Technical Support: 1-800-424-6723 • www.bio-rad.com 7Fig. 1. Schematic representation of an adsorption isotherm. A, operating line describing the

8 Technical Support: 1-800-424-6723 • www.bio-rad.comNote: 1. The slope of the operating line is equal to the phase ratio (β). 2. The feed concent

Technical Support: 1-800-424-6723 • www.bio-rad.com 9The time interval used for sample addition dictates the number of time points. At the end of

10 Technical Support: 1-800-424-6723 • www.bio-rad.com2.7 Design of Experiments Design of experiments (DoE) is a systematic investigation of the e

Technical Support: 1-800-424-6723 • www.bio-rad.com 11Section 3High-Throughput Experiments3.1 Manual WorkflowA manual workflow for HT experimentat

12 Technical Support: 1-800-424-6723 • www.bio-rad.com3.2 Types of Studies3.2.1 Screening To utilize the full potential of HT experimentation, s

Technical Support: 1-800-424-6723 • www.bio-rad.com 133.2.2 Adsorption Isotherm An adsorption isotherm can be determined using a ladder of initi

14 Technical Support: 1-800-424-6723 • www.bio-rad.comCsalt (M) …C0 (mg/mL) 1 2 3 … 10 11 12C1AC2BC3CC4DC5EC6FC7GC8H(A)Csalt1Csalt4C0 (mg/mL) C1C2C3

Technical Support: 1-800-424-6723 • www.bio-rad.com 15Csalt (M) …C0 (mg/mL) 1 2 3 … 10 11 12C1AC2BC3CC4DC5EC6FC7GC8H(A)Csalt1Csalt4C0 (mg/mL) C1C2

16 Technical Support: 1-800-424-6723 • www.bio-rad.comLoading Order first lasttime (min)t1t2t3…t10t11t12C0 (mg/mL) 1 2 3 … 10 11 12ABCDEFGΔt = tfina

Technical Support: 1-800-424-6723 • www.bio-rad.com 173.3 General Considerations3.3.1 SampleThe sample should be clarified using a 0.45 or 0.22

18 Technical Support: 1-800-424-6723 • www.bio-rad.comFor instance, compared to a buffer solution, a protein solution of relatively high protein con

Technical Support: 1-800-424-6723 • www.bio-rad.com 193.4 Reliability of the HT experimentation Data: Error Analysis and Error Propa

20 Technical Support: 1-800-424-6723 • www.bio-rad.comCorrPODCm= Corr22Corr21PC OD mODmmσσ σ  = +−    Where Cp and σp represent the calc

Technical Support: 1-800-424-6723 • www.bio-rad.com 210204060801001201401601800.00 2.00 4.00 6.00 8.00 10.00q (mg/mL_media)c (mg/mL)DataLangmuir m

22 Technical Support: 1-800-424-6723 • www.bio-rad.com3.5 Equipment and Materials Required for HT ExperimentsIn addition to the Foresight

Technical Support: 1-800-424-6723 • www.bio-rad.com 23Recommended settings for vacuum filtration are 1 min at –1” Hg. If liquid is still visible i

24 Technical Support: 1-800-424-6723 • www.bio-rad.com3.6.3 Sample Loadinga) Volume and Concentration• Load 300 μl of sample per well, cover the

ContentsSection 1 Introduction 1Section 2 Theoretical Background 2

Technical Support: 1-800-424-6723 • www.bio-rad.com 253.6.4 Washing Weakly Bound Material• Add 300 μl of wash buffer per well and cover the pla

26 Technical Support: 1-800-424-6723 • www.bio-rad.comSection 4Application Example 4.1 IgG isotherm on cPrime™ Chromatographic MediaIntrod

Technical Support: 1-800-424-6723 • www.bio-rad.com 27Materials and Methods1. Preparation of stock solutions:Loading buffer: 0.02 M Na Acetate, p

28 Technical Support: 1-800-424-6723 • www.bio-rad.comc) Collect the supernatant from each well by centrifugation (300 x g, 3 min) onto a 96-well c

Technical Support: 1-800-424-6723 • www.bio-rad.com 29Fig. 6. HT experimentation Isotherm data for hIgG on Nuvia™ cPrime™ media, at pH of 4.5 and

30 Technical Support: 1-800-424-6723 • www.bio-rad.comSection 5Ordering Information5.1 Foresight™ Chromatography Media Filter Plates Product Mode

Technical Support: 1-800-424-6723 • www.bio-rad.com 315.2 Foresight Chromatography Media ColumnsProduct Mode Catalog #Foresight Nuvia S Column,

32 Technical Support: 1-800-424-6723 • www.bio-rad.com5.3 Foresight™ RoboColumns® UnitsProduct Mode Catalog #Foresight™ Nuvia™ S, RoboColumn, 200

Technical Support: 1-800-424-6723 • www.bio-rad.com 33Section 6References1. Bensch M et al. (2005). High throughput screening of chromatographic

Section 4 Application Example 264.1 IgG isotherm on Bio-Rad’s cPrime™ Chromatographic Media

Life Science Group Sig 121110024413 Rev A US/EGBio-Rad Laboratories, Inc. Web site www.bio-rad.com USA 800 424 6723 Australia 61 2 9914 280

Technical Support: 1-800-424-6723 • www.bio-rad.com 1Section 1IntroductionThe advent of high-throughput (HT) experimentation has revolutionized th

2 Technical Support: 1-800-424-6723 • www.bio-rad.comSection 2Theoretical Background2.1 Batch AdsorptionBatch adsorption occurs when a liquid phas

Technical Support: 1-800-424-6723 • www.bio-rad.com 3Normally, V0 is much larger than the latter two liquid volumes, hence the assumption that V =

4 Technical Support: 1-800-424-6723 • www.bio-rad.comTo use Eq. 3a, only the flowthrough concentration must be measured. Conversely, using Eq. 3c req