Size Exclusion BioHPLC Columns
|
|
- Lynn James
- 6 years ago
- Views:
Transcription
1 Size Exclusion BioHPLC Columns Size Exclusion Product Families Particle Porosity Functionalities Particle Pore Size Application Sizes Agilent Bio SEC- Silica Fully porous N/A um 00A, 0A, 00A High efficiency Silica Fully porous N/A um 00A, 0A, 00A More pore sizes 00A, 000A, 000A ProSEC 00S Silica Fully porous N/A um 00A Extend linear range broader sample range ZORBAX GFC Silica Fully porous N/A um, 6um 0A 00A Larger column sizes. High efficiency small particles. Multiple pore sizes to cover broad range of molecular sizes. Single pore size to cover range of globular proteins/antibodies Page 6. Larger columns for fractionation
2 SEC Applications Non-Denaturing Conditions Separations Based on Size in Solution Impurity testing, aggregation g analysis (separation of monomer/dimer/aggregates) Molecular weight characterization over wide MW range (000 0M) possible Expression and folding studies Separation of Reaction Components and Products, (conjugates) Purification Desalting and salt exchange Collection of fractions under non-denaturing conditions Page 6 SEC Technique Technique is isocratic No interaction with the stationary phase Separation based on differences in size Eluent contains buffer and salt Technique does not denature proteins 0,000 µv 00,000 0,000 00,000 0,000 00,000 0,000 00,000 0,000 00,000 0,000 Decreasing size R&D Bio SEC.DATA Smaller molecules spend longer in the pores and elute later. Larger molecules spend less time in the pores and elute sooner. 0 RT [min] 0 Gel Filtration Standard ( vial diluted with mls of eluent) (Thyroglobulin, -globulin, ovalbumin, myoglobin & vitamin B)
3 Elution Profiles If sample molecules have the same molecular dimensions, they will coelute and may not be separated by this technique If two molecules have the same molecular weight but different size in solution they may be separated The calibration curve describes how different size molecules elute fromthecolumnandcanbeusedto determine molecular weight equivalents based on solution size Choose the Appropriate Pore Size Pore Size MW Range Typical Sample (Globular Protein) 00Å 00 00,000 Small proteins 0Å 00 0,000 Small proteins 00Å,000,0,000 Most protein samples 00Å,000,000,000 Larger proteins and antibodies 000Å 0,000 7,00,000 Very large proteins 000Å > 0,000,000 Very large molecules viruses Lower limit on this column is hard to define, Use only for special applications 6
4 Example of Choice of Pore Size Data. Thyroglobulin Aggregates-. Thyroglobulin-MW 670KD.. -globulin- MW 8 KD. Ovalbumin- MW KD 6. Myoglobin- MW 7KD 7. Vitamin B-MW.KD Eluent: Columns: Flow: Detector: System: 0mM Na HPO, 0mM NaH PO + 0.M NaCl, ph6.8 Agilent Bio SEC- 00Å, 0Å & 00Å µm.6x00mm 0.ml/min UV@0nm Agilent 60 Infinity Bio-Inert LC System. Dimer.. Monomer... Monomer Fragment.. Azide. Retained Molecule Bio-Rad Gel Filtration Standards Mix 6 7 Mouse IgG 00Å 0Å 00Å 0Å 00Å 00Å 7 Separation of Recombinant EPO-Protein (MW: kda) Agilent BioSEC-, 00A.6 x 00mm Column: Agilent Bio SEC, 00A.6 x 00mm ID Sample: repo protein (0.mg/ml) Buffer: 0mM Na phosphate, ph 7.0 Flow rate: 0.ml/min Temp: C Gradient: isocratic Detection: UV Baseline expansion
5 Monitoring Aggregation and Impurities (IgG MW; 0,000 Da) BioSEC- 00Å, Phosphate buffer to control ph and provide ionic strength Optimized to eliminate interactions 0.0 Column: Bio SEC- 00Å, 7.8x00mm Mobile phase: 0 mm Phosphate, ph Flow rate:.0 ml/min Temperature: Ambient 0.0 Sample: Monoclonal antibody (0 L, mg/ml) MAb Aggregates MAb Dimer Low Molecular Weight Impurities Buffer/Excipients utes 9 Match Particle Size to Flow Rate and Column Pressure Pressure Increasing flow rate reduces efficiency and resolution.0ml/min.ml/min.0 ml/min monomer dimer Flow Rate Particle Size Column: Agilent Bio SEC-, 7.8 x0mm Sample: mab (mg/ml) Injection: ul Flow rate:.0,. and ml/min (6 bar, 7 bar, 0 bar) Eluent: 0mM sodium phosphate + 00mM Na-sulfate Detection: 0nm Flow Rate Resolution Monomer/Dimer Monomer Efficiency Percentage Dimer.0ml/min.,0 0.6.ml/min., utes.0ml/min.,
6 Agilent NEW Size Exclusion Columns Agilent Bio SEC- Unique, m particle 00Å, 0Å, 00Å pore sizes Highest resolution Highest efficiency Faster SEC separations m Particle 00Å, 0Å, 00Å, 00Å, 000Å, 000Å pore sizes High stability and long lifetime Great reproducibility Comparing Agilent Bio SEC- and Bio SEC- Improved Efficiency With Smaller Particles Agilent Bio SEC-, 00Å, 7.8x00mm Column Pressure Bio SEC- (7.8x00mm) 9 bar Bio SEC- (7.8x00mm) bar Peak Protein SEC-, 00Å (7.8x00mm) SEC-, 00Å (7.8x00mm) Thyroglobulin 60 0 BSA Dimer BSA Ribonuclease A Uracil , 00Å, 7.8x00mm Column: Bio SEC- 00Å and Bio SEC- 00Å Buffer: 0 mm Phosphate buffer, ph 7 Flow rate:.0 ml/min for 7.8x00 mm Temperature: Ambient (~ C) Detection: UV nm Injection: 0 µl ( L for.6x00 mm) Sample: ) Thyroglobulin (.0 mg/ml), 670 kd; ) BSA dimer, kd; ) BSA (.0 mg/ml), 66 kd; ) Ribonuclease A (.0 mg/ml),.7 kd, and ) Uracil (. g/ml), 0D. 6
7 Agilent Bio SEC- Sharp Peaks and High Resolution of Proteins Agilent Bio SEC-, 0Å, 7.8x00mm Agilent Bio SEC-, 00Å, 7.8x00mm Peak Compound SEC- 0Å SEC- 00Å Thyroglobulin BSA Dimer 0 0 BSA Ribonuclease A Columns: Bio SEC-, 7.8x00 mm Mobile phase: 0 mm Phosphate, ph 7 Flow rate:.0 ml/min Temperature: Ambient (~ C) Detection: UV nm Injection: 0 µl Sample: ) Thyroglobulin (.0 mg/ml), 670 kd ) BSA dimer, kd ) BSA (.0 mg/ml), 66 kd ) Ribonuclease A (.0 mg/ml),.7 kd ) Uracil (. g/ml), 0D Uracil Agilent Bio SEC- Bio SEC-, 0Å, Capacity: BSA Loading Test.00 Injection volume: 0 L BSA Concentration.80 (from low to high): mg/ml mg/ml mg/ml.0 0 mg/ml.0 0 mg/ml 0 mg/ml Column: Bio SEC-, 0Å, 7.8x00mm Mobile phase: 0 mm Phosphate, ph 7.0 Flow Rate:.0 ml/min Injection volume: 0 µl Detector: nm
8 Agilent Bio SEC- Separations of E. coli lysate Bio SEC- 0Å, 7.8x00mm Bio SEC- 00Å, 7.8x00mm Column: Column: Bio SEC- 0Å, 7.8x00mm & Bio SEC- 00Å, 7.8x00mm Mobile phase: 0. M Phosphate, ph 7.0 Flow rate:.0 ml/min Detection: UV nm Temperature: Ambient Injection: 0 µl Sample: E. coli lysate (. mg/ml) Agilent Bio SEC- Monoclonal Antibody Aggregation Monitoring 0.0 Column: Bio SEC- 00Å, 7.8x00mm Mobile phase: 0 mm Phosphate, ph Flow rate:.0 ml/min Temperature: Ambient 0.0 Sample: Monoclonal antibody (0 L, mg/ml) utes 8
9 High Capacity and High Resolution Au nm m Å. thyroglobulin,. min. BSA dimer, 6.9 min. BSA monomer, 6.9 min. ribonuclease A, 8.7 min. poly-dl-alanine (- kda), 9.90 min 6. uracil,. min Time (min) nm m TSK G000SW xl. thyroglobulin,.6 min. BSA dimer, 6. min. BSA monomer, 7.0 min. ribonuclease A, 9. min. poly-dl-alanine (- kda), 0.0 min 6. uracil,.8 min Time (min) High Capacity and High Resolution Columns: Bio SEC-,.6x00 mm Buffer: 0. M Phosphate buffer, ph 7.0 Flow rate: 0. ml/min Detector: UV nm Injection volume: 0 µl.thyroglobulin. -Globulin. Ovalbumin. Ribonuclease A. p-aminobenzoic acid Bio SEC-, 0Å Bio SEC-, 00Å Bio SEC-, 00Å
10 High Molecular Weight Protiens 0. Column: Bio SEC-, 000Å,.6x00mm Buffer:0 mm phosphate buffer, ph Flow rate: 0. ml/min Detection: nm 0.8 Temperature: Ambient Proteins:.Thyroglobulin (670kD). Uracil (0D) Possible Dimer Reproducibility, 00Å, 7.8x00mm Buffer: 0 mm Phosphate buffer, ph 7.0 Flow Rate:.0 ml/min Detector: nm Injection: 0 ul.thyroglobulin. BSA. Ribonuclease A. Uracil utes 0
11 Lot to Lot Reproducibility, 00Å, 7.8x00mm Black: SN 086 Lot# 00907MGB Blue: SN 0 Lot# 0606MG Green: SN 7 Lot# 006CL utes Lot Thyroglobulin BSA Ribonuclease A Uracil RT Eff. RT Eff. RT Eff. RT Eff MG MG CL Stability at ph 8.. Thyroglobulin. BSA. Ribonuclease A. Uracil Bio SEC-, 00Å,.6x00mm Buffer: 0mM Phosphate, ph 8. Wavelength: nm Flow Rate: 0. ml/min Injection: ul Pre-test, ph7.0 Day AM, ph8. Day PM, ph8. Day AM, ph8. Day PM, ph8. Day AM, ph8. Day 7 AM, ph utes
12 Stability at ph 8., increasing injection volume.00 Column: Bio SEC-, 00Å, 7.8x00mm, ph Sample: BSA Concentration: mg/ml Mobile Phase: 0 mm Phosphate, ph 8..0 Flow Rate:.0 ml/min Detector: utes Injection volumes ul, 0 ul, 0 ul, 0 ul, and 00 ul Monoclonal Antibody Aggregation Monitoring Columns: Bio SEC-, 00Å, 7.8x00mm Buffer: 0 mm Phosphate, ph 7 Flow Rate:.0 ml/min Sample: Mab Temperature: Ambient MAb Dimer Mab Aggregates Buffer Peak
13 Size Exclusion Separations of E. coli Lysate (Single Columns) Columns: Bio SEC-, 7.8x00mm Buffer: 0 mm Phosphate, ph 7.0 Flow Rate:.0 ml/min Detector: nm Injection: 0 µl (. mg/ml) SEC-, 000Å SEC-, 00Å SEC-, 00Å SEC-, 0Å Size Exclusion Separations of E. coli Lysate (Columns In Series)...0 Columns: Bio SEC-, 000Å, 7.8x00mm + Bio SEC-, 0Å, 7.8x00mm Buffer: 0 mm Phosphate, ph 7.0 Flow Rate:.0 ml/min Detector: t nm Injection: 0 µl (. mg/ml) utes
BioHPLC columns. Tim Rice Biocolumn Technical Specialist
BioHPLC columns Tim Rice Biocolumn Technical Specialist AU Typical Application Areas Size Exclusion: Aggregation Analysis Ion Exchange: Charge Isoform Analysis 0.035 Monomer 0.030 0.025 0.020 0.015 Dimer
More informationAgilent AdvanceBio SEC Columns for Aggregate Analysis: Instrument Compatibility
Agilent AdvanceBio SEC Columns for Aggregate Analysis: Instrument Compatibility Technical Overview Introduction Agilent AdvanceBio SEC columns are a new family of size exclusion chromatography (SEC) columns
More informationProSEC 300S. Protein Characterization columns
ProSEC 300S Protein Characterization columns Agilent s ProSEC 300S is a silica-based material specifically designed for the analysis of proteins by aqueous size exclusion chromatography. With a proprietary
More informationR R Innovation Way P/N SECKIT-7830 Newark, DE 19711, USA Tel: Fax: Website: Published in November 2013
5-100 Innovation Way Newark, DE 19711, USA Tel:302-3661101 Fax:302-3661151 Website: www.sepax-tech.com Published in November 2013 P/N SECKIT-7830 These Phases are developed based on innovative surface
More informationSeparate and Quantify Rituximab Aggregates and Fragments with High-Resolution SEC
Separate and Quantify Rituximab Aggregates and Fragments with High-Resolution SEC The Agilent 126 Infinity Bio-Inert Quaternary LC System and the AdvanceBio SEC 3Å, 2.7 µm Column Application Note Biologics
More informationPhysical Stability of a Silica- Based Size Exclusion Column for Antibody Analysis
Physical Stability of a Silica- Based Size Exclusion Column for Antibody Analysis Atis Chakrabarti* and Roy Eksteen + Tosoh Bioscience LLC, King of Prussia, PA 19406 *Corresponding Author. + Current address:
More information2012 Waters Corporation 1
UPLC User meeeting April 2012 Principles and Practices for SEC, IEX for Intact Protein Analysis by UPLC anders_feldthus@waters.com 2012 Waters Corporation 1 Agenda Ion-Exchange Chromatography Theory and
More informationAnalysis of biomolecules by SEC and Ion-Exchange UPLC
Analysis of biomolecules by SEC and Ion-Exchange UPLC Anders Feldthus, Waters Nordic 2011 Waters Corporation 1 Waters Commitment To develop, commercialize and market columns that when used on Waters ACQUITY
More informationHigh-throughput and Sensitive Size Exclusion Chromatography (SEC) of Biologics Using Agilent AdvanceBio SEC Columns
High-throughput and Sensitive Size Exclusion Chromatography (SEC) of Biologics Using Agilent AdvanceBio SEC Columns Agilent AdvanceBio SEC 3 Å, 2.7 µm columns Application note Bio-Pharmaceutical Author
More informationChoosing the Right Calibration for the Agilent Bio SEC-3 Column
Choosing the Right Calibration for the Agilent Bio SEC-3 Column Application Note Biotherapeutics & Biosimilars Author Keeley Mapp Agilent Technologies, Inc Introduction Size exclusion chromatography (SEC)
More informationADVANCE ACCURACY AND PRODUCTIVITY FOR FASTER ANALYSIS
Agilent AdvanceBio Columns ADVANCE ACCURACY AND PRODUCTIVITY FOR FASTER ANALYSIS with Agilent ZORBAX RRHD 3Å 1.8 µm columns ns Rapid resolution high definition columns for UHPLC protein and peptide separations
More informationNext Generation Zirconia-Based Antibody Purification Media
Next Generation Zirconia-Based Antibody Purification Media Dr. Clayton McNeff, Dwight Stoll, Danielle Hawker (ZirChrom), Dr. Andy Clausen (Merck) Dr. Peter W. Carr and Dr. Anuradha Subramanian (U of MN)
More informationSuperdex 200 Increase columns
GE Healthcare Life Sciences Data file 29-452-69 AA Size exclusion chromatography Superdex 2 Increase columns Superdex 2 Increase prepacked columns (Fig 1) are designed for size exclusion chromatography
More informationProtein Quantitation using various Modes of High Performance Liquid Chromatography
Platzhalter für Bild, Bild auf Titelfolie hinter das Logo einsetzen Protein Quantitation using various Modes of High Performance Liquid Chromatography Grotefend, S.; Wroblewitz, S.; Kaminski, L.; Limberger,
More information'Tips and Tricks' for Biopharmaceutical Characterization using SEC
'Tips and Tricks' for Biopharmaceutical Characterization using SEC Waters Corporation 2012 Waters Corporation 1 Waters Commitment To develop, commercialize and market columns that, when used on Waters
More informationHIC as a Complementary, Confirmatory Tool to SEC for the Analysis of mab Aggregates
HIC as a Complementary, Confirmatory Tool to SEC for the Analysis of mab Aggregates Julia Baek, Shanhua Lin, Xiaodong Liu, Thermo Fisher Scientific, Sunnyvale, CA Application Note 2126 Key Words Size exclusion
More informationSeparation of Recombinant Human Erythropoietin (repo) Using Agilent Bio SEC-3
Separation of Recombinant Human Erythropoietin (repo) Using Agilent Bio SEC-3 Application Note BioPharma Authors Phu T Duong and James Martosella Agilent Technologies, Inc. 285 Centerville Rd, Wilmington,
More informationApplication Note. Author. Abstract. Biopharmaceuticals. Verified for Agilent 1260 Infinity II LC Bio-inert System. Sonja Schneider
Combining small-scale purification and analysis of monoclonal antibodies on one instrument Protein purification with high-volume injection using the Agilent 126 Infinity Bio-inert Quaternary LC System
More informationSize Exclusion Chromatography
Size Exclusion TSKgel PW-type PW PW XL TOSOH BIOSCIENCE TOSOH BIOSCIENCE LLC 156 Keystone Drive Montgomeryville, PA 18936-9637 Orders & Service: (800)366-4875 Fax: (215)283-5035 www.tosohbioscience.com
More informationMolecular Characterization of Biotherapeutics The Agilent 1260 Infi nity Multi-Detector Bio-SEC Solution with Advanced Light Scattering Detection
Molecular Characterization of Biotherapeutics The Agilent 126 Infi nity Multi-Detector Bio-SEC Solution with Advanced Light Scattering Detection Application Note Biologics and Biosimilars Authors Sonja
More informationAnalysis of Intact and C-terminal Digested IgG1 on an Agilent Bio MAb 5 µm Column
Analysis of Intact and C-terminal Digested IgG1 on an Agilent Bio MAb µm Column Application Note BioPharma Authors Xiaomi Xu and Phu T Duong Agilent Technologies, Inc. Abstract Nearly all proteins undergo
More informationmabs and ADCs analysis by RP
mabs and ADCs analysis by RP Shanhua Lin, Ph.D. The world leader in serving science Protein and mab Separation by HPLC Size difference? YES Size Exclusion Chromatography (SEC) MAbPac SEC-1 NO NO Charge
More informationLifetime stability of size exclusion chromatography columns for protein aggregate analysis
APPLICATION NOTE 72362 Lifetime stability of size exclusion chromatography columns for protein aggregate analysis Authors Amy Farrell 1, Craig Jakes 1, Alexander Ley 2, Mauro De Pra 2, Frank Steiner 2,
More informationCharacterization of mab aggregation using a Cary 60 UV-Vis Spectrophotometer and the Agilent 1260 Infinity LC system
Characterization of mab aggregation using a Cary 60 UV-Vis Spectrophotometer and the Agilent 1260 Infinity LC system Application Note Biopharmaceuticals Authors Arunkumar Padmanaban and Sreelakshmy Menon
More informationBio-Monolith Protein G Column - More Options for mab Titer Determination
Bio-Monolith Protein G Column - More Options for mab Titer Determination Application Note Biologics and Biosimilars Author Phu T. Duong Agilent Technologies, Inc. Introduction In recent years, monoclonal
More informationSize Exclusion Chromatography of Biosimilar and Innovator Insulin Using the Agilent AdvanceBio SEC column
Size Exclusion Chromatography of Biosimilar and Innovator Insulin Using the Agilent AdvanceBio SEC column Application Note Bio-Pharmaceutical Authors M. Sundaram Palaniswamy and Andrew Coffey Agilent Technologies,
More informationCapillary Electrophoresis of Proteins
Capillary Electrophoresis of Proteins SDS Capillary Gel Electrophoresis SDS-CGE Outline CE-SDS Gel Analysis Description of Technique Method Development Tips PA800 plus kits SDS-MW IgG Purity & Heterogeneity
More informationA universal chromatography method for aggregate analysis of monoclonal antibodies
APPLICATION NOTE A universal chromatography method for aggregate analysis of monoclonal antibodies No. 2161 Amy Farrell 1, Jonathan Bones 1, and Ken Cook 2 1 NIBRT, Dublin, Ireland; 2 Thermo Fisher Scientific,
More informationGel Filtration Calibration Kits
GE Healthcare Life Sciences Gel Filtration Calibration Kits Product booklet Codes: 28-4038-41 Low Molecular Weight 28-4038-42 High Molecular Weight Page finder 1. Legal 3 2. Handling 4 2.1. Safety warnings
More informationmab Titer Analysis with the Agilent Bio-Monolith Protein A Column
mab Titer Analysis with the Agilent Bio-Monolith Protein A Column Application Note Biopharmaceuticals and Biosimilars Authors Emmie Dumont, Isabel Vandenheede, Pat Sandra, and Koen Sandra Research Institute
More informationPromix TM. Enter a New Era in Biomolecule Analysis with. Columns. Unsurpassed Selectivity and Peak Capacity for Peptides and Proteins
Promix TM Enter a New Era in Biomolecule Analysis with Promix TM Columns Unsurpassed electivity and Peak Capacity for Peptides and Proteins Applications: Proteomics Peptide/Protein Analysis Peptide/Protein
More informationAZURA Bio LC. 2 FPLC and Purification Solutions
AZURA Bio LC 2 FPLC and Purification Solutions Flexible and adjustable to your biopurification tasks Fast protein liquid chromatography (FPLC) of bio molecules can be a task. KNAUER offers with AZURA an
More informationIon Exchange Chromatography Media. Cellufine IEX. Technical Data Sheet
Ion Exchange Chromatography Media IEX Technical Data Sheet 2-1, Otemachi 2-Chome Chiyoda-ku, Tokyo 1-815 JAPAN Phone +81-3-3243-615 / Fax +81-3-3243-6219 e-mail: cellufine@jnc-corp.co.jp web: http: //www.jnc-corp.co.jp/fine/en/cellufine/
More informationFast mass transfer Fast separations High throughput and improved productivity Long column lifetime Outstanding reproducibility Low carryover
columns ProSwift Reversed-Phase Monolith Columns for Protein Analysis ProSwift reversed-phase columns use a unique monolith technology for fast, high-resolution HPLC and LC/MS separations of proteins.
More informationDisulfide Linkage Analysis of IgG1 using an Agilent 1260 Infinity Bio inert LC System with an Agilent ZORBAX RRHD Diphenyl sub 2 µm Column
Disulfide Linkage Analysis of IgG1 using an Agilent 126 Infinity Bio inert LC System with an Agilent ZORBAX RRHD Diphenyl sub 2 µm Column Application Note Biotherapeutics & Biosimilars Author M. Sundaram
More informationDevelopment of Analysis Methods for Therapeutic Monoclonal Antibodies Using Innovative Superficially Porous Particle Biocolumns
Development of Analysis Methods for Therapeutic Monoclonal Antibodies Using Innovative Superficially Porous Particle Biocolumns Anne Blackwell Bio Columns Product Support Scientist Suresh Babu Senior Application
More informationSuperdex 75 Increase columns
GE Healthcare Size exclusion chromatography Superdex 7 Increase columns Superdex 7 Increase prepacked columns are designed for rapid separation and analysis of proteins with molecular weights ranging from
More informationTSKgel STAT Columns for High Performance Ion Exchange Chromatography
Separation Report No. 09 TSKgel STAT Columns for High Performance Ion Exchange Chromatography Table of Contents. Introduction. Basic Characteristics of TSKgel STAT Columns - Characteristics of Packing
More informationACQUITY UPLC Protein BEH SEC Columns and Standards
and Standards CONTENTS I. INTRODUCTION II. CONFIGURING AN ACQUITY UPLC SYSTEM FOR USE IN SEC PROTEIN SEPARATIONS a. Calibrators III. GETTING STARTED a. ecord installation b. Column connectors c. Column
More informationFast and High Resolution Analysis of Intact and Reduced Therapeutic Monoclonal Antibodies (mabs)
Fast and High Resolution nalysis of Intact and Reduced Therapeutic Monoclonal ntibodies (mbs) The gilent io-inert L and dvanceio RP-mb olumns pplication Note io-pharmaceutical uthor M. Sundaram Palaniswamy
More information3. Close the bottom end of the column and apply the packing device on top. Pump water through the upper adaptor to remove air.
INSTRUCTIONS FOR USE WorkBeads Protein A Product name Pack size Article number WorkBeads Protein A Bulk Media 1.5 ml 40605001 Bulk Media 5 ml 40605002 Bulk Media 10 ml 40605003 Bulk Media 100 ml 40605004
More informationUserguide: SepFast Gel Filtration columns
Userguide: SepFast Gel Filtration columns 1. Introduction Gel Filtration is a proven technique which is widely used for size-based molecular separation. SepFast Gel Filtration resin is specially designed
More informationINSECT CELL/BACULOVIRUS PRODUCTION
INSECT CELL/BACULOVIRUS PRODUCTION PEF # GENE NAME TRANSFER VECTOR BEVS MOLECULAR WEIGHT 2015-XXXX XXXX pbac1 flashbacultra TM 36.0 kda EXPRESSION METHOD OVERVIEW: Insect cells Spodoptera frugiperda (Sf9)
More informationSize-Exclusion Chromatography (SEC) Optimization Guide
Size-Exclusion Chromatography (SEC) Optimization Guide 1 Table of Contents I. Introduction II. ACQUITY UPLC Instrumentation Considerations for Successful SEC Analysis III. Importance of Developing a Robust
More informationPRINCIPLES OF CHROMATOGRAPHY SEC COLUMNS
COLUMNS PRINCIPLES OF CHROMATOGRAPHY The analysis, isolation, and purifi cation of biomolecules can be accomplished by a number of chromatographic modes. Each mode is based on specific physical, chemical,
More informationHigh Resolution Analysis of Heparin and Heparin-like Impurities on Glycomix TM an Anion Exchange Column
High Resolution Analysis of and -like Impurities on Glycomix TM an Anion Exchange Column Authors Xiaomi Xu Yuna Li Aiyan Cui Haiying Chen Sepax Technologies, Inc. 5 Innovation Way Newark, DE 19711 USA
More informationMAbPac SCX-10 Columns
User Manual MAbPac SCX-10 Columns 065393 Revision 09 October 2015 For Research Use Only. Not for use in diagnostic procedures. Product Manual for MAbPac SCX-10, 10µm Analytical Column (10μm, 9 x 250 mm,
More informationAn effective platform for purification of IgM monoclonal antibodies using Hydroxyapatite
An effective platform for purification of IgM monoclonal antibodies using Hydroxyapatite Frank Hensel, Patrys, GmbH Pete Gagnon, Validated Biosystems 5th International Conference on Hydroxyapatite and
More informationPolishing of monoclonal antibodies using Capto S ImpAct
Application note 29-83-27 AA Ion exchange chromatography Polishing of monoclonal antibodies using Capto S ImpAct Capto S ImpAct chromatography medium (resin) is a strong cation exchanger (CIEX). The medium
More informationApplication of Agilent AdvanceBio Desalting-RP Cartridges for LC/MS Analysis of mabs A One- and Two-dimensional LC/MS Study
Application of Agilent AdvanceBio Desalting-RP Cartridges for LC/MS Analysis of mabs A One- and Two-dimensional LC/MS Study Application note Biotherapeutics and Biologics Authors Suresh Babu C.V., Anne
More informationProtein-Pak Hi Res HIC Column and HIC Protein Standard
Protein-Pak Hi Res HIC Column and HIC Protein Standard CONTENTS I. INTRODUCTION II. a. Mobile Phase b. Flow Direction CONNECTING COLUMN TO LC SYSTEM I. INTRODUCTION This offering contains non-porous, polymethacrylate-based
More informationTechnical Data. Analytical Data
Technical Data High resolution analysis of monoclonal antibodies utilizing cation exchange column YMC-BioPro SP-F Analysis of Monoclonal Antibody (MAb) Pharmaceuticals Using Non-Porous Type Ion Exchange
More informationAgilent BioHPLC Columns PROTEIN IDENTIFICATION AND IMPURITY PROFILING USING REVERSED-PHASE HPLC/UHPLC
Agilent BioHPLC Columns PROTEIN IDENTIFICATION AND IMPURITY PROFILING USING REVERSED-PHASE HPLC/UHPLC REVERSED-PHASE HPLC/UHPLC Agilent can boost your accuracy and productivity Reversed-phase is used to
More informationEcono-Pac High Capacity Ion Exchange Cartridges, 5 ml Instruction Manual. Catalog Numbers , ,
Econo-Pac High Capacity Ion Exchange Cartridges, 5 ml Instruction Manual Catalog Numbers 732-0026, 732-0027 732-0066, 732-0067 Table of Contents Section 1 Introduction... 1 Section 2 Connection to Bio-Rad
More informationZwitterion Chromatography ZIC
Zwitterion Chromatography ZIC A novel technique, with unique selectivity, suitable for preparative scale separations? PhD Einar Pontén What is Zwitterion Chromatography? Our definition: Liquid chromatography
More informationTypes of chromatography
Chromatography Physical separation method based on the differential migration of analytes in a mobile phase as they move along a stationary phase. Mechanisms of Separation: Partitioning Adsorption Exclusion
More informationA Fast and Robust Linear ph Gradient Separation Platform for Monoclonal Antibody (MAb) Charge Variant Analysis
A Fast and Robust Linear ph Gradient Separation Platform for Monoclonal Antibody (MAb) Charge Variant Analysis Shanhua Lin, Julia Baek, and Chris Pohl Thermo Fisher Scientific, Sunnyvale, CA, USA Overview
More informationCell Clone Selection Using the Agilent Bio-Monolith Protein A Column and LC/MS
Cell Clone Selection Using the Agilent Bio-Monolith Protein A Column and LC/MS Application Note Biopharmaceuticals and Biosimilars Authors Emmie Dumont, Isabel Vandenheede, Pat Sandra, Koen Sandra Research
More informationSepax Technologies, Inc.
Sepax Technologies, Inc. Sepax Technologies, Inc. develops and manufactures products in the area of chemical and biological separations, bio-surfaces and proteomics. Sepax product portfolio includes 1)
More informationBioPro IEX Resins. Resin Downstream Purification
BioPro IEX Resins Resin Downstream Purification 2 BioPro: Designed for bioseparations Contents 3 Introduction...4 Guide to IEX Media Selection...5 Specifications...7 Solutions for Capture Processes...9
More informationMolecules are separated by size. Small molecules enter pores of stationary phase (b), large molecules do not (a) Larger molecules elute first
Molecules are separated by size Small molecules enter pores of stationary phase (b), large molecules do not (a) Larger molecules elute first Types of gels for open column, preparative scale GPC (i) Bio-Gel
More informationHiLoad Superdex 30 prep grade HiLoad Superdex 75 prep grade HiLoad Superdex 200 prep grade
GE Healthcare Life Sciences Data file 8--52 AE Gel filtration HiLoad Superdex 3 prep grade HiLoad Superdex 75 prep grade HiLoad Superdex 2 prep grade HiLoad columns are XK laboratory columns prepacked
More informationTSKgel Size Exclusion Chromatography Columns
TSKgel Size Exclusion Chromatography Columns Silica-based for protein analysis: TSKgel SW mab TSKgel SW TSKgel SWXL TSKgel SuperSW Polymer-based for desalting: TSKgel BioAssist DS Columns Polymethacrylate-based
More informationIgG Purity/Heterogeneity and SDS-MW Assays with High- Speed Separation Method and High Throughput Tray Setup
IgG Purity/Heterogeneity and SDS-MW Assays with High- Speed Separation Method and High Throughput Tray Setup High Throughput Methods to Maximize the Use of the PA 800 Plus system Jose-Luis Gallegos-Perez
More informationBivalirudin Purification:
Bivalirudin Purification: Sorbent Screening and Overload Experiments Marc Jacob, Joshua Heng, and Tivadar Farkas Phenomenex, Inc., 411 Madrid Ave., Torrance, CA 90501 USA PO94190412_W Abstract In this
More informationNuvia S and Q High-Capacity Ion Exchange Media Instruction Manual
... Nuvia S and Q High-Capacity Ion Exchange Media Instruction Manual Catalog numbers 156-0311 156-0411 156-0313 156-0413 156-0315 156-0415 156-0317 156-0417 Please read these instructions before you use
More informationAnalysis of amoxicillin and five impurities on the Agilent 1220 Infinity LC System
Analysis of amoxicillin and five impurities on the Agilent Infinity LC System LC analysis of impurities down to the.% level with long sub--µm columns, high flow rates and back pressure greater than bar
More informationApplication Note. Biopharm. Author. Abstract. Christian Wenz Agilent Technologies, Inc. Waldbronn, Germany
Performance of commercially available gels for protein characterization by capillary gel electrophoresis with UV detection on the Agilent 7100 CE System Application Note Biopharm Author Christian Wenz
More informationApplication Note USD Purification of Mouse IgM from Cell Culture Supernatant by Cation Exchange Chromatography on CM Ceramic HyperD F Sorbent
Application Note USD 241 Purification of Mouse IgM from Cell Culture Supernatant by Cation Exchange Chromatography on CM Ceramic HyperD F Sorbent What this Study Demonstrates T h i s s t u d y o n C a
More informationThe practical task of monoclonal IgG purification with CHT TM ceramic hydroxyapatite
The practical task of monoclonal IgG purification with CHT TM ceramic hydroxyapatite Pete Gagnon, Jie He, Paul Ng, Julia Zhen, Cheryl Aberin, Heather Mekosh 11th Annual Waterside Conference, Chicago, May
More informationFundamentals and Techniques of Preparative HPLC. Parto Zist Behboud Tel: 42108
Fundamentals and Techniques of Preparative HPLC Parto Zist Behboud www.partoz.com Tel: 42108 Today s Overview 1 Basic know-how for up-scaling 2 Practical examples of up-scaling 3 Recycle preparation 4
More informationFast and Efficient Peptide Mapping of a Monoclonal Antibody (mab): UHPLC Performance with Superficially Porous Particles
Fast and Efficient Peptide Mapping of a Monoclonal Antibody (mab): UHPLC Performance with Superficially Porous Particles Application Note Biotherapeutics and Biosimilars Authors James Martosella, Alex
More informationMethod Development Considerations for Reversed-Phase Protein Separations
Method Development Considerations for Reversed-Phase Protein Separations Hillary B. Hewitson, Thomas E. Wheat, Paula Hong, Kenneth J. Fountain APPLICATION BENEFITS n The BEH00 C 4 chemistry is available
More informationGel filtration columns and media
GE Healthcare Life Sciences Gel filtration columns and media Selection guide General information Principles of gel filtration Gel filtration (GF) separates molecules on the basis of differences in size
More informationModule 16: Gel filtration: Principle, Methodology & applications. Dr. Savita Yadav Professor Department of Biophysics AIIMS, New Delhi
PAPER 9: TECHNIQUES USED IN MOLECULAR BIOPHYSICS I Module 16: Gel filtration: Principle, Methodology & applications Dr. Savita Yadav Professor Department of Biophysics AIIMS, New Delhi Module 16 Gel filtration:
More informationG Sep Size Exclusion Columns
G-Biosciences 1-800-628-7730 1-314-991-6034 technical@gbiosciences.com A Geno Technology, Inc. (USA) brand name G Sep Size Exclusion Columns (Cat. # 786 1297, 786 1298, 786 1300, 786 1301, 786 1302, 786
More informationFast and High-Resolution Reversed-Phase Separation of Synthetic Oligonucleotides
Fast and High-Resolution Reversed-Phase Separation of Synthetic Oligonucleotides High-pH-stable, superficially porous particle columns for LC/UV and LC/MS Application Note Biologics and Biosimilars Authors
More informationCapture of mouse monoclonal antibodies
Capture of mouse monoclonal antibodies by cation exchange chromatography Life Science Products Processing Capture of mouse monoclonal antibodies from cell culture supernatant by cation exchange chromatography
More informationGel Filtration Chromatography. Teaching Kit Manual. GeNei TM. Cat No. New Cat No. KT Revision No.:
Gel Filtration Chromatography Teaching Kit Manual Cat No. New Cat No. KT39 106190 Revision No.: 00130405 CONTENTS Page No. Objective 3 Principle 3 Kit Description 6 Materials Provided 7 Procedure 8 Observation
More informationDeveloping Robust and Efficient IEX Methods for Charge Variant Analysis of Biotherapeutics Using ACQUITY UPLC H-Class System and Auto Blend Plus
Developing Robust and Efficient IEX Methods for Charge Variant Analysis of Biotherapeutics Using ACQUITY UPLC H-Class System and Auto Blend Plus Robert Birdsall, Thomas Wheat, and Weibin Chen Waters Corporation,
More informationAnalytical Instrument Qualification According to USP <1058>: Requirements and Examples for the Agilent 1290 Infinity LC System
Analytical Instrument Qualification According to USP : Requirements and Examples for the Agilent 1290 Infinity LC System 1) USP : Scope, approach, requirements 2) Qualification Examples for
More informationFraction Analysis of Cysteine Linked Antibody-Drug Conjugates Using Hydrophobic Interaction. chromatography. Agilent 1260 Infinity II Bio-Inert System
Application Note Biologics & Biosimilars Fraction Analysis of Cysteine Linked Antibody-Drug Conjugates Using Hydrophobic Interaction Chromatography Agilent 126 Infinity II Bio-Inert System 7 6 5 4 5. 7.5
More informationThe Eshmuno Chromatography Family of Resins
The Eshmuno Chromatography Family of Resins The perfect accessory for highly productive downstream purification EMD Millipore is a division of Merck KGaA, Damstadt, Germany Introduction For over 300 years,
More informationAffi-Gel Protein A MAPS II Kit Instruction Manual
Affi-Gel Protein A MAPS II Kit Instruction Manual Catalog Number 153-6159 For Technical Service Call Your Local Bio-Rad Office or in the U.S. Call 1-800-4BIORAD (1-800-424-6723) Table of Contents Introduction...1
More informationPractical Issues in the Industrial use of Hydroxyapatite for Purification of Monoclonal Antibodies
Practical Issues in the Industrial use of Hydroxyapatite for Purification of Monoclonal Antibodies Pete Gagnon, Validated Biosystems Inc., 240 Avenida Vista Montana Suite 7F, San Clemente, CA 92672 Paul
More informationNPTEL VIDEO COURSE PROTEOMICS PROF. SANJEEVA SRIVASTAVA
LECTURE-06 PROTEIN PURIFICATION AND PEPTIDE ISOLATION USING CHROMATOGRAPHY TRANSCRIPT Welcome to the proteomics course. Today, we will talk about protein purification and peptide isolation using chromatography
More informationrprotein A GraviTrap Protein G GraviTrap rprotein A/Protein G GraviTrap
GE Healthcare Life Sciences Data file 28-9921-04 AA rprotein A GraviTrap Protein G GraviTrap rprotein A/Protein G GraviTrap Protein sample preparation rprotein A GraviTrap, Protein G GraviTrap, and rprotein
More informationFlexibility with. Preparative HPLC
Flexibility with Preparative HPLC Date: August 26 th, 2014 Time: 09.00 10.00 am CEST 1 Who are your Presenters? Dr. Christian Textor Product Manager Preparative HPLC Dipl. Biol. Hagen Schlicke Product
More informationElectronic Supplementary Information for The effect of protein concentration on the viscosity of a recombinant albumin solution formulation
Electronic Supplementary Material (ESI) for RSC Advances. This journal is The Royal Society of Chemistry 2016 Electronic Supplementary Information for The effect of protein concentration on the viscosity
More informationGenotoxicity is the property of a compound
Impurities Analysis in Pharmaceuticals: Genotoxicity is the property of a compound known to have irreversible effects on the structure and functionality of the DNA in cells and cause DNA loss, DNA replication
More informationCombining High Temperature and Small Particles: The Advantages of Zirconia
Combining High Temperature and Small Particles: The Advantages of Zirconia EAS 9 Dan owlan 1, Bingwen Yan 1, Clayton V. Mceff 1, R.A. Henry 1 ZirChrom Separations, Inc. 617 Pierce St., Anoka, M 5533 Independent
More informationMaximizing Chromatographic Resolution of Peptide Maps using UPLC with Tandem Columns
Maximizing Chromatographic Resolution of Peptide Maps using UPLC with Tandem Columns Hongwei Xie, Martin Gilar, and Jeff Mazzeo Waters Corporation, Milford, MA U.S. APPLICATION BENEFITS The ACQUITY UPLC
More informationelectrophoresis tech Performance Comparison of the Experion Automated Electrophoresis System and SDS-PAGE for Protein Analysis
electrophoresis tech note 5299 Performance Comparison of the Experion Automated Electrophoresis System and for Protein Analysis Karen Zhu, Marie Nguyen, William Strong, and Christina Whitman-Guliaev, Bio-Rad
More informationAnaTag HiLyte Fluor 647 Protein Labeling Kit
AnaTag HiLyte Fluor 647 Protein Labeling Kit Catalog # 72049 Kit Size 3 Conjugation Reactions This kit is optimized to conjugate HiLyte Fluor 647 SE to proteins (e.g., IgG). It provides ample materials
More informationApplication Note. Author. Abstract. Biotherapeutics and Biologics. Sonja Schneider Agilent Technologies, Inc. Waldbronn, Germany
D-LC/MS Characterization of Charge Variants Using Ion Exchange and Reversed-Phase Chromatography Multiple Heart-Cutting D-LC Analysis of Innovator versus Biosimilar Monoclonal Antibodies Application Note
More informationImproved Peptide Maps Using Core-Shell Media: Explaining Better Biopharmaceutical Applications With Kinetex C18 Columns
Improved Peptide Maps Using Core-Shell Media: Explaining Better Biopharmaceutical Applications With Kinetex C18 Columns Michael McGinley, Jeff Layne, and Vita Knudson Phenomenex, Inc., 411 Madrid Ave.,Torrance,
More informationPeptide Mapping: A Quality by Design (QbD) Approach
Peptide Mapping: A Quality by Design (QbD) Approach Application Note Bio-Pharmaceutical Authors Sreelakshmy Menon and Suresh babu C.V. Agilent Technologies, Inc. Richard Verseput S-Matrix Corporation Abstract
More information1. Bloomsbury BBSRC Centre for Structural Biology, Birkbeck College and University College London.
Purification/Polishing of His-tagged proteins - Application of Centrifugal Vivapure Ion-exchange Membrane Devices to the Purification/Polishing of Histagged Background Multi-milligram quantities of highly
More informationMATF Antigen Submission Details and Standard Project Deliverables
MATF Antigen Submission Details and Standard Project Deliverables What we require when you submit your antigen: Proteins For a recombinant protein target, we require a minimum of 400µg soluble recombinant
More information