Monoclonal antibodies
|
|
- Francine Montgomery
- 8 years ago
- Views:
Transcription
1 MAb Contaminant Removal with a Multimodal Anion Exchanger A Platform Step to Follow Protein A Kjell Eriksson, Anders Ljunglöf, Gustav Rodrigo, and Eggert Brekkan Reprinted with permission from BioProcess International 7(2) (February 2009) Monoclonal antibodies (MAbs) constitute ~30% of the biopharmaceutical products currently under development (1). An increasing demand for MAbs during the past decade has led to intense development of highexpression cell cultures (2). Today, it is possible to see titers of 4 5 g/l, and expression levels as high as 15 g/l and greater have been reported. As a consequence, demand has increased for more efficient downstream processes. That demand, combined with its potential for reducing time-to-market, has increased interest in the value of platform approaches to MAb production (upstream as well as downstream). Many MAb producers are now adopting such approaches. Today, nearly all approved MAb processes include a capture step using protein A. In generic protocols with Pr o d u c t Fo c u s : Monoclonal antibodies Pr o c e s s Fo c u s : Downstream processing Wh o Sh o u l d Re a d: Process development, manufacturing Ke y w o r d s : HCP, multimodal chromatography, platform technologies, aggregates Le v e l: Intermediate direct capture of MAb from clarified cell culture supernatant, the high selectivity of protein A resins provides a high yield of very pure product. Protein A thus forms the foundation for the success of most MAb platform approaches to downstream processing. With the recently introduced Capto adhere multimodal anion-exchange resin, which has high selectivity for contaminants that remain after protein A separation, an additional step has been taken toward developing highly effective MAb downstream processes (3). The high purity that follows capture on protein A resins, combined with the multimodal functionality of Capto adhere resin, provides for highly productive, two-step processes (4, 5). MAb Platform Toolbox A platform process includes a number of unit operations, methods, and conditions based on experience gained from purifying a class of molecules that have similar properties ( 10). With >20 approved products and >10 in clinical trials, MAbs are just such a class of molecule (1). And the cornerstone for all MAb platforms is the unique selectivity of protein A. A platform technology enables rapid and economical process development and scale-up. By facilitating shorter development times, it potentially helps a greater number of drug candidates pass through development laboratories. The GE Healthcare ( use of highly similar platform approaches can also lead to greater robustness and smoother process transfer from development to manufacturing. Because MAbs are similar but not identical, their platforms cannot be completely fixed in all details; they must be flexible. The chromatography steps that follow a protein A step might, therefore, shift in order or be slightly different from process to process. A toolbox concept (Figure 1) could therefore be used to establish an efficient MAb purification process. For optimal results, it is most 52 BioProcess International February 2009
2 Figure 1: MAb chromatography resin toolbox *WO 2004/45, patent application from Lonza, may be relevant to a process that includes protein A followed by anion-exchange chromatography. Figures 1 GE Healthcare, reproduced with permission. Cell Culture Cell Removal Figure 2: The Capto adhere ligand n-benzyln-methyl-ethanolamine has the potential for several types of interactions, the most pronounced being ion interaction, hydrogen bonding, and hydrophobic interaction. OH OH O O N + OH MabSelect SuRe Virus Inactivation and Filtration SP Sepharose FF/Capto S Pool for Final Filtration Ultrafiltration/Diafiltration important to optimize every step in a MAb process, including capture with protein A. Our suggested platform includes MabSelect SuRe protein A resin for capture, although MAbSelect and MabSelect Xtra brands also work very well for MAb platforms. The MabSelect SuRe product contains an alkalistabilized, protein A derived ligand that can be used with M NaOH as a clean-in-place (CIP) agent, thereby prolonging its working life over that of other protein A resins (11, 12). The ligand is more stable in regards to proteolysis than a regular (native or recombinant) protein A ligand, which further reduces ligand leakage. In addition, its lack of Fab binding allows for more generic elution conditions such as (13) for a broad range of MAbs and Fc fusion proteins. All that facilitates the platform 0.2 µm Sterile Filtration Capto Q approach. The unique selectivity achievable with protein A media such as MabSelect SuRe brand makes a twochromatography step process possible. Such processes have been reported (14, 15). A Mu l t i m o d a l Anion Exchanger GE Healthcare s Capto adhere anion exchanger offers multimodal functionality designed for post protein A polishing in MAb processes. Contaminants left in a product pool after protein A capture are removed by operating Capto adhere resin in flowthrough mode so that they bind to the resin. Figure 2 shows the Capto adhere ligand: n-benzyl-n-methyl ethanolamine. This ligand displays several different modes of interaction, the most dominant being ionic interaction. However, other types of interaction such as hydrogen bonding and hydrophobic interaction also contribute. Multimodal anion exchangers have different selectivities from those of traditional strong anion exchangers (e.g., Q-types). Figure 3 compares descending gradient elution in bind elute mode of five different MAbs from Capto adhere and Capto Q resins. The order of elution is the same on both, but the at which the different MAbs elute is considerably lower with the Capto adhere product. This indicates that it involves additional interactions and that the multimodal functionality results in different selectivity. Capto adhere resin was initially designed for MAb purification, and its successful use in purification of an Fc-fusion protein has recently been reported (1). Nevertheless, this does not exclude using the multimodal anion exchanger in other applications such as the purification of other types of recombinant proteins. Op t i m i z at i o n o f Co n d i t i o n s Capto adhere resin is used following protein A in MAb purification processes. Its use to remove remaining contaminants is preferably done in flowthrough mode under conditions that Table 1: Overview of contaminant removal that can be expected with Capto adhere resin following protein A capture (MVM = minute virus of mice; MuLV = murine leukemia virus; LRV = log reduction factor) Contaminant/Step Protein A Eluate LRF: Capto adhere Contaminant Clearance HCP ,000 ppm 2 3 Protein A 5 10 ppm 2 3 DNA 10 1,000 ppb 3 4 Viruses MVM MuLV N/A Aggregated MAb 1 20% 1 2 Table 2: Virus removal efficiency of the Capto adhere multimodal anion exchanger; minute virus of mice (MVM) and murine leukemia virus (MuLV) were applied in flow-through mode with IgG MAb at.75 (LRF = log reduction factor at 95% confidence limit). Virus Conductivity (ms/cm) LRF MVM 10 >5. MVM 30 >5.9 MuLV ±0.4 MuLV 30 3.±0.4 Fe b r u a r y 2009 BioProcess International 53
3 Figure 3: Comparing elution of five different antibodies on Capto adhere and Capto Q resins. Antibodies elute in the same order on both but much earlier (at a higher ) on Capto Q. The two were eluted using different gradient ranges ( for Capto adhere resin and for Capto Q resin) MAb 4:.22 MAb 3: 5.25 MAb 2: 5.12 MAb 1: 4.93 MAb 5: ml allow antibodies to pass directly through a column while contaminants are adsorbed. In flow-through mode, loading conditions will be a compromise between those favoring yield and those favoring contaminant clearance. So loading conditions need to be optimized, and there will be a trade-off between yield and purity. Optimization is preferably done using design of experiments (DoE). When optimizing Capto adhere loading conditions, we have found that load,, and conductivity are the most Capto Q MAb 3, 9.5 MAb 4, 9. MAb 5, 9.41 MAb 2,.37 MAb 1, ml important factors. Optimizing them will influence both yield and clearance of contaminants. In DoE, all factors are varied simultaneously in a structured approach. A common method is to define a reference experiment (center point) and then perform other experiments around that point. DoE Example: The starting material was clarified cell culture supernatant containing monoclonal IgG1. Protein A chromatography used for the capture step was followed by viral inactivation at low. Polishing was then performed Figure 5: Example of response surfaces for yield and host-cell protein (HCP), dimers/aggregates (D/A), and leached protein A clearance Yield Conductivity 15 ms/cm Host Cell Protein Load 300 mg/ml Figure 4: Full factorial design for optimizing load conditions (sample load varied mg/ml,, and conductivity 2 15 ms/cm) 2 75 Load Cond. on Capto adhere resin in flow-through mode. A full-factorial design in three variables (load,, and conductivity) with three center points was set up to investigate the effects on yield and clearance of key contaminants: host cell proteins (HCP), dimers/aggregates (D/A), and leached protein A (Figure 4). Separate models were created for yield, clearance of HCP, D/A, and leached protein A (Figure 5). The response surfaces show how load,, and conductivity influence the different responses and how to reach desired values for each of them. Sweet-spot analysis was performed with the following criteria for acceptance: With a load 100 g/l resin, yield in flowthrough should be 90%, HCP 50 ppm, D/A 1%, and leached protein A 5 ppm. The sweet-spot (Figure ) is a point (red) at which the goals for yield and contaminant clearance are fulfilled. Because the goals for HCP and protein A removal are fulfilled in the entire experimental space, the optimization in this particular example was done between yield and D/A removal. Dimer/Aggregates Conductivity 15 ms/cm Protein A Load 300 mg/ml Figure : Sweet-spot analysis including all experimental data from the experimental design Load BioProcess International Fe b r u a r y 2009
4 Op e r at i o n Capto adhere separation is operated so that antibodies pass directly through a column while contaminants are adsorbed. One contaminant that needs to be reduced/removed is the MAb itself in aggregated form. Using Capto adhere resin reduces not only the amount of aggregates, but also other contaminants of concern such as HCP, DNA, protein A, and viruses. Table 1 overviews the post protein A contaminant removal that can be expected using this multimodal anion exchanger. It is of particular interest to establish virus-removal efficiency. A study was thus performed to investigate removal of minute virus of mice (MVM) and murine leukemia virus (MuLV). Table 2 shows the results. Both model viruses are effectively removed at higher and lower ionic strengths. For a traditional anion exchanger such as a Q-resin, good virus clearance would not be expected at the higher ionic strength. Combining a protein A resin s high selectivity with the unique contaminantremoving properties of the Capto adhere anion exchanger makes it possible to cut many purification processes from three chromatography steps to two. To achieve this in practice, it is of utmost importance to optimize not only the multimodal anion-exchange step, but also the initial capture step. The newly developed MabSelect SuRe resin, with its alkaline-stable ligand that allows CIP with NaOH, is an obvious choice for this purpose. Key advantages are its proven long working life, high dynamic binding capacity, and low ligand leakage (11, 12, 17, 1). To further optimize the capture step, in some cases it may be necessary to develop an intermediate wash step that removes loosely bound contaminants before elution (19). Figure 7 shows one example of a two-step purification process for a MAb. The starting material was clarified Chinese hamster ovary (CHO) cell supernatant spiked with host cell proteins (HCPs). Capture was performed using MabSelect SuRe protein A resin. After sample application, an intermediate wash was run with 20 mm sodium phosphate, 5% isopropanol, and 0.5 M NaCl at 7.0. Analysis of the wash fraction showed that it mainly contained aggregates and HCP (19). MAbs were then eluted with sodium citrate 3.4. The HCP concentration was reduced from 130,000 ppm to 55 ppm, and the remaining aggregate content was 0.7%. MabSelect SuRe eluate was then purified further in flowthrough mode on Capto adhere anionexchange resin. As can be seen from the Table 3: In support of Figure 7 (HCP = host cell protein; D/A = dimers/aggregates; ND = not determined) Two-Step Process Yield (%) D/A (%) Protein A (ppm) HCP (ppm) Start material spiked with HCP ,000 MabSelect SuRe <1 55 Capto adhere 95 <0.1 ND 7.5 results, contaminant removal was excellent. HCP levels were reduced to <10 ppm, and both aggregate and leached protein A levels were below detection limits. A two-step chromatography process offers obvious advantages over a threestep process: Fewer steps improve yield and help shorten process times, translating to higher productivities (in grams produced over time). A two-step platform also improves process economy. In addition, less chromatography resin is used, less equipment and buffer volumes are needed, and the required floor space in a production facility can be reduced. When working on process optimization from productivity as well as process economy viewpoints, following a LEAN concept is highly advantageous. Reducing waste, materials, and especially time will lead to more efficient production (20). As described here, Capto adhere resin is usually run in flow-through mode, but in some cases it could be advantageous to use it in bind elute mode (1). In addition, some MAb preparations contain fragment levels that are too high for the final product. MAb fragments bind less strongly to this resin and could thus be separated from monomeric MAbs. Dimers and other aggregates bind more strongly and thereby elute after monomeric MAbs. Running this multimodal anion exchanger in bind elute mode thus has some advantages in certain situations. But it also has one major drawback: loading capacity. Loading in flow- Figure 7: Purification of a MAb from CHO cell culture (MabSelect SuRe capture step followed by Capto adhere resin in flow-through mode) A 20 nm (mau) 4,000 MAbSelect SuRe 10 A 20 nm (mau) 3,000 ms/cm 150 3,000 2, ,000 1, ,000 ml 4 1, ml Fe b r u a r y 2009 BioProcess International 55
5 through mode is often g/l resin; in bind elute mode it will be in the range g/l. However, the possibility of using Capto adhere resin in bind elute mode should make it an interesting alternative for cases in which its unique selectivity is required and also for proteins other than MAbs. Re f e r e n c e s 1 Roque ACA, et al. Antibodies and Genetically Engineered Related Molecules: Production and Purification. Biotechnol. Prog. 20(3) 2004: Birch JR, Racher AJ. Antibody Production. Adv. Drug Deliv. Rev. 5, 200: Johansson B-L, et al. Preparation and Characterization of Prototypes for Multi- Modal Separation Aimed for Capture of Positively Charged Biomolecules at High-Salt Conditions. J. Chromatogr. A 101(1) 2003: Johansson HJ, et al. Multi-Modal Chromatography for Purification of Monoclonal Antibodies (poster). Recovery of Biological Products XII 2 7 April 200, (Phoenix, AZ). American Chemical Society Biotechnology Division; www. recoveryconferences.org. 5 Eriksson K, et al. Post Protein A Removal of Contaminants from Monoclonal Antibodies with a Multi-Modal Anion Exchanger. 234th ACS National Meeting August 2007 (Boston, MA). American Chemical Society; Slaff G. Application of Technology Platforms to the Purification of Monoclonal Antibodies. BioProcess International European Conference April 2005 (Berlin, Germany). IBC Life Sciences, com. 7 Sofer G, Chirica LC. Improving Productivity in Downstream Processing. BioPharm Int. 19, 200: Shukla AA, et al. Downstream Processing of Monoclonal Antibodies: Application of Platform Approaches. J. Chromatogr. B 4, 2007: Ishihara T, Kadoya T. Accelerated Purification Process Development of Monoclonal Antibodies for Shortening Time to Clinic: Design and Case Study of Chromatography Processes. J. Chromatogr. A 117 (1 2) 2007: Low D, et al. Future of Antibody Purification. J. Chromatogr. B 4, 2007: Hahn R. Comparison of Protein A Affinity Sorbents III: Life Time Study. J. Chromatogr. A 1102, 200: Hober S, et al. Protein A Chromatography for Antibody Purification. J. Chromatogr. B 4, 2007: Ghose G, et al. Antibody Variable Region Interactions with Protein A: Implications for the Development of Generic Purification Processes. Biotechnol. Bioeng. 92() 2005: Vunnum S, et al. Anion Exchange Purification of Monoclonal Antibodies: Principles of Weak Partitioning Chromatography. 232nd ACS National Meeting September 200 (San Francisco, CA); American Chemical Society, 15 Kelley B. Very Large Scale Monoclonal Antibody Purification: The Case for Conventional Unit Operations. Biotechnol. Prog. 23(5) 2007: Rea DW, et al. Solutions for Purification of Fc-fusion Proteins. BioPharm Int. supplement, March 200: Johansson HJ. Advances in the Purification of Monoclonal Antibody Based Therapeutics. Antibody Development and Production 1 3 March 200 (Carlsbad, CA); IBC Life Sciences, 1 Jagschies G, et al. Technical and Economical Evaluation of Downstream Processing Options for Monoclonal Antibody (MAb) Production. BioPharm Int. supplement, June 200: Grönberg A, et al. A Strategy for Development of a MAb Purification Platform (poster). BioProcess International European Conference February 200 (Prague, Czech Republic); IBC Life Sciences, www. informa-ls.com. Also BioProcess Int. 5(1) 2007: Jagschies G, et al. Modern Tools and LEAN Concepts for Manufacturing Flexibility. BioPharm Int. (in press). c Corresponding author Kjell Eriksson is a senior scientist in Life Sciences R&D at GE Healthcare Bio-Sciences AB, Björkgatan 30, SE Uppsala, Sweden; , kjell.eriksson@ge.com; www. gelifesciences.com. Anders Ljunglöf, Gustav Rodrigo, and Eggert Brekkan are all scientists at GE Healthcare. Capto, MabSelect, MabSelect SuRe, MabSelect Xtra and Sepharose are trademarks of GE Healthcare companies. Figures GE Healthcare, reproduced with permission. 5 BioProcess International Fe b r u a r y 2009
Process-scale purification of monoclonal antibodies polishing using Capto Q
GE Healthcare Life Sciences Application note 28-937-16 AB Ion exchange chromatography Process-scale purification of monoclonal antibodies polishing using Capto Q Summary Anionic exchange media are an industry
More informationContinuous Chromatography for Monoclonal Antibody Purification from Cell Culture Supernatant
Continuous Chromatography for Monoclonal Antibody Purification from Cell Culture Supernatant Massimo Morbidelli Institute for Chemical and Bioengineering, ETH Zurich, Switzerland www.morbidelli.ethz.ch
More informationApplication Note. Separation of three monoclonal antibody variants using MCSGP. Summary
Application Note Separation of three monoclonal antibody variants using MCSGP Category Matrix Method Keywords Analytes ID Continuous chromatography, Biochromatography; FPLC Protein A-purified monoclonal
More informationMonoclonal Antibody Production: Building the Platform. Andrew Clutterbuck Eden Biodesign Ltd.
Monoclonal Antibody Production: Building the Platform Andrew Clutterbuck Eden Biodesign Ltd. Questions Questions are encouraged throughout the presentation and can be asked by using the email address provided
More informationA novel AIEX chromatography medium (resin) to remove IgA and IVIG purification process
A novel AIEX chromatography medium (resin) to remove IgA and IVIG purification Guodong Javier Jia 1, Chor Sing Tan 2, Linus Laurin 3, Henrik Ihre 3, Lili Sui 1 1 Fast Trak China, GE Healthcare Life Sciences,
More informationApplication Note. Purifying common light-chain bispecific antibodies using MCSGP. Summary
Application Note Purifying common light-chain bispecific antibodies using MCSGP Category Matrix Method Keywords Analytes ID Continuous chromatography, biochromatography Antibodies MCSGP Bispecific antibody,
More informationApplication Note. Increasing the activity of monoclonal antibody isoforms by MCSGP. Summary
Application Note Increasing the activity of monoclonal antibody isoforms by MCSGP Category Matrix Method Keywords Countercurrent chromatography, FPLC Antibodies MCSGP FPLC, Biobetters, MCSGP, countercurrent
More informationBringing Downstream Productivity into Phase with Upstream Antibody Production
Bringing Downstream Productivity into Phase with Upstream Antibody Production Pete Gagnon, Validated Biosystems 3 rd International Monolith Symposium, Portoroz, May 30 June 4, 2008 The need for speed When
More informationPOROS CaptureSelect affinity columns for highspeed quantification of IgG Fc fusion proteins
APPLICATION NOTE POROS CaptureSelect affinity chromatography columns POROS CaptureSelect affinity columns for highspeed quantification of IgG Fc fusion proteins Introduction POROS columns, containing highperformance
More informationEvaluation of an alkali stable Protein A matrix versus Protein A Sepharose Fast Flow and considerations on process scale-up to 20,000L.
Evaluation of an alkali stable Protein A matrix versus Protein A Sepharose Fast Flow and considerations on process scale-up to 20,000L. Simon C.B. Jones & Martin P. Smith. LONZA, 224 Bath Road, Slough,
More informationMethod Development for Size-Exclusion Chromatography of Monoclonal Antibodies and Higher Order Aggregates
Method Development for Size-Exclusion Chromatography of Monoclonal Antibodies and Higher Order Aggregates Paula Hong and Kenneth J. Fountain Waters Corporation, 34 Maple St., Milford, MA, USA APPLICATION
More informationHow To Test For Cleaning Efficiency With A Predictor 96 Well Filter Plate
GE Healthcare Life Sciences Application note 28-9845-64 AA Process chromatography High-throughput process development for design of cleaning-in-place protocols Cleaning-in-place (CIP) of chromatography
More informationHigh-throughput Process Development with PreDictor Plates
GE Healthcare High-throughput Process Development with PreDictor Plates Principles and Methods imagination at work Handbooks from GE Healthcare GST Gene Fusion System Handbook 18-1157-58 Affinity Chromatography
More informationAffi-Prep Protein A Matrix Instruction Manual
Affi-Prep Protein A Matrix Instruction Manual Catalog Numbers 156-0005 156-0006 Bio-Rad Laboratories, 2000 Alfred Nobel Dr., Hercules, CA 94547 LIT-230 Rev B Table of Contents Section 1 Introduction...1
More informationChromatographic Methods for the Purification of Monoclonal Antibodies and their Alternatives:A Review
Chromatographic Methods for the Purification of Monoclonal Antibodies and their Alternatives:A Review Ishan Arora 1 1 Department of Chemical Engineering, Indian Institute of Technology Delhi Abstract Monoclonal
More informationThe market for therapeutic monoclonal antibodies
PURIFYING THERAPEUTIC MONOCLONAL ANTIBODIES Amit Mehta, Martha Lovato Tse, Jace Fogle, Amy Len, Roshan Shrestha, Nuno Fontes, Bénédicte Lebreton, Bradley Wolk, Robert van Reis Genentech Eliminating the
More informationChallenges in the cgmp Manufacturing of hescs: Lessons Learned from Monoclonal Antibodies
Challenges in the cgmp Manufacturing of hescs: Lessons Learned from Monoclonal Antibodies ISCT 2011 Annual Meeting Rotterdam, The Netherlands May 18 21, 2011 BioProcess Technology Consultants www.bptc.com
More informationA new, integrated, continuous purification process template for monoclonal antibodies
A new, integrated, continuous purification process template for monoclonal antibodies Alex Xenopoulos* Alison Dupont, Christopher Gillespie, Ajish Potty, Michael Phillips Processing Technologies Merck
More informationRESOURCE Q, 1 ml and 6 ml RESOURCE S, 1 ml and 6 ml
GE Healthcare Life Sciences Instructions 71-7146-00 AI Ion Exchange Columns RESOURCE Q, 1 ml and 6 ml RESOURCE S, 1 ml and 6 ml Introduction RESOURCE Q and S are pre-packed columns for separating biomolecules
More informationGain efficiency in your process development with ÄKTA avant
Gain efficiency in your process development with ÄKTA avant gelifesciences.com Gain efficiency in your process development with ÄKTA avant Development of efficient manufacturing processes is a necessity
More informationManufacturing processes for
Single-Use, Continuous- Countercurrent, Multicolumn Chromatography Marc Bisschops, Lynne Frick, Scott Fulton, and Tom Ransohoff Reprinted with permission from BioProcess International 7(6) (June 2009)
More informationKMS-Specialist & Customized Biosimilar Service
KMS-Specialist & Customized Biosimilar Service 1. Polyclonal Antibody Development Service KMS offering a variety of Polyclonal Antibody Services to fit your research and production needs. we develop polyclonal
More informationEden Biodesign ebook Monoclonal Antibody Production: Building the Platform
Eden Biodesign ebook Monoclonal Antibody Production: Building the Platform CHAPTER 1: Overview CHAPTER 2: Challenges CHAPTER 3: Purification Methodology CHAPTER 4: Results CHAPTER 5: About Eden Biodesign
More informationRecent advances in the purification of IgM monoclonal antibodies
Recent advances in the purification of IgM monoclonal antibodies Pete Gagnon, Validated Biosystems Frank Hensel, Paul Andrews, Patrys, Ltd. Richard Richieri, Avid BioServices, Inc. 3 rd Wilbio Conference
More informationMustang Q XT Chromatography Capsules
USD 2599 Mustang Q XT Chromatography Capsules High Throughput, Scalable, and Reusable Ion Exchange Membrane Chromatography Meeting Process Demands for Scalability and Economy The use of ion exchange chromatography
More informationIntroduction to Bioprocessing
Introduction to Bioprocessing Cambridge Healthtech Institute Peptalk Palm Springs, CA Presented by Susan Dana Jones and Sheila Magil BioProcess Technology Consultants www.bptc.com BioProcess Technology
More informationBiopharmaceutical Process Evaluated for Viral Clearance
Authored by S. Steve Zhou, Ph.D. Microbac Laboratories, Inc., Microbiotest Division The purpose of Viral Clearance evaluation is to assess the capability of a manufacturing production process to inactivate
More informationGuide to Reverse Phase SpinColumns Chromatography for Sample Prep
Guide to Reverse Phase SpinColumns Chromatography for Sample Prep www.harvardapparatus.com Contents Introduction...2-3 Modes of Separation...4-6 Spin Column Efficiency...7-8 Fast Protein Analysis...9 Specifications...10
More informationTECHNICAL BULLETIN. HIS-Select Nickel Affinity Gel. Catalog Number P6611 Storage Temperature 2 8 C
HIS-Select Nickel Affinity Gel Catalog Number P6611 Storage Temperature 2 8 C TECHNICAL BULLETIN Product Description HIS-Select Nickel Affinity Gel is an immobilized metalion affinity chromatography (IMAC)
More informationMabSelect SuRe. GE Healthcare Life Sciences
GE Healthcare Life Sciences Instructions 11-0026-01 AD Affinity Chromatography MabSelect SuRe MabSelect SuRe is an alkali-tolerant protein A-derived medium for capturing monoclonal antibodies from large
More informationAdvances in Biopharmaceutical and Vaccine Manufacturing Plants
Hitachi Review Vol. 62 (2013), No. 4 267 Advances in Biopharmaceutical and Vaccine Manufacturing Plants Sei Murakami, Dr. Eng. Haruo Suzuki Keisuke Shibuya, Dr. Sc. OVERVIEW: The development of innovative
More informationCustomer Application Brief. Filtration Processes Applied in Therapeutic Monoclonal Antibody Production. Bioprocess, Biologicals, & Pharmaceutical
Customer Application Brief Bioprocess, Biologicals, & Pharmaceutical Filtration Processes Applied in Therapeutic Monoclonal Antibody Production Introduction Monoclonal antibodies were among the first biotechnology
More informationHis GraviTrap. GE Healthcare. Operation
GE Healthcare Data File 11-0036-90 AB Affinity purification His GraviTrap His GraviTrap is a prepacked, single-use column for purification of histidine-tagged proteins by immobilized metal affinity chromatography
More informationDuring regulatory inspections,
B I O P R O C E S S TECHNICAL Cleaning and Cleaning Validation in Process Chromatography Current Industry Practices and Future Prospects Gail Sofer and Jonathan Yourkin During regulatory inspections, manufacturers
More informationLFB GROUP & SANOFI combine their bioproduction capabilities to provide integrated CMO services for biopharmaceuticals
LFB GROUP & SANOFI combine their bioproduction capabilities to provide integrated CMO services for biopharmaceuticals MAbLaunch TM is a joint bioproduction platform combining LFB Biomanufacturing (LFB
More informationGuidance for Industry. Monoclonal Antibodies Used as Reagents in Drug Manufacturing
Guidance for Industry Monoclonal Antibodies Used as Reagents in Drug Manufacturing U.S. Department of Health and Human Services Food and Drug Administration enter for Drug Evaluation and Research (DER)
More informationVirus Purification with Membrane Chromatography
Virus Purification with Membrane Chromatography 1 st Workshop European Network of Viral Vaccines Processes, October 14-15, 2010, Frankfurt am Main Stefan Fischer-Frühholz, Laura Chirica, Miyako Hirai,
More informationBiotechpharma company profile
Biotechpharma company profile October 2013 1 History 2004 Biotechpharma UAB established as a proteomic research company in Vilnius, Lithuania 2005 Company became a member of UK s Northway group, investing
More informationTHE DEVELOPMENT OF THERAPEUTIC MONOCLONAL ANTIBODY PRODUCTS
THE DEVELOPMENT OF THERAPEUTIC MONOCLONAL ANTIBODY PRODUCTS Table of Contents CHAPTER 1: Overview of Monoclonal Antibody Therapeutics CHAPTER 2: Monoclonal Antibody Discovery Technologies CHAPTER 3: CMC
More informationPOROS CaptureSelect affinity columns for rapid, small-scale purification and sample preparation of recombinant proteins
APPLICATION NOTE POROS CaptureSelect affinity chromatography columns POROS CaptureSelect affinity columns for rapid, small-scale purification and sample preparation of recombinant proteins Introduction
More informationChoose your optimal tools for protein studies
Protein Purification Choose your optimal tools for protein studies Bacterial Baculoviral Cell free Mammalian Secreted Intracellular High yield Increased solubility Highest purity Highest yield His-tag
More informationProtein Purification Handbook
Protein Purification Handbook Protein Purification Handbook 18-1132-29 Edition AC Handbooks from Amersham Biosciences Antibody Purification Handbook 18-1037-46 The Recombinant Protein Handbook Protein
More informationTechnology Transfer of CMC Activities for MAb Manufacturing. 2010 ge healthcare (www.gelifesciences.com)
M a n u f a c t u r i n g OPERATIONS Technology Transfer of CMC Activities for MAb Manufacturing by Patricia Seymour, Susan Dana Jones, Howard L. Levine With combined 2009 revenues estimated to be over
More informationApplication Note. USD 2995 (a) High Throughput Regeneration Study on MEP HyperCel Mixed-Mode Sorbent on ScreenExpert RoboColumns u
Application Note USD 2995 (a) High Throughput Regeneration Study on MEP HyperCel Mixed-Mode Sorbent on ScreenExpert RoboColumns u Summary A regeneration study of MEP HyperCel mixed-mode sorbent was conducted
More informationHiTrap Heparin HP, 1 ml and 5 ml
Instructions 71-7004-00 AU HiTrap affinity columns HiTrap Heparin HP, 1 ml and 5 ml HiTrap Heparin HP is a prepacked ready to use, column for preparative affinity chromatography. The special design of
More informationFast Trak Training & Education
GE Healthcare Life Sciences Fast Trak Training & Education Our expertise, your advantage imagination at work Contents General course information General course information 3 Upstream processing Advanced
More informationEfficient Multi-Well Protein Purification Strategies
Application Note PN 33576 Efficient Multi-Well Protein Purification Strategies Introduction Many tools and techniques are available today for protein purification. Development of a purification process
More informationClassic Immunoprecipitation
292PR 01 G-Biosciences 1-800-628-7730 1-314-991-6034 technical@gbiosciences.com A Geno Technology, Inc. (USA) brand name Classic Immunoprecipitation Utilizes Protein A/G Agarose for Antibody Binding (Cat.
More informationTransgenic technology in the production of therapeutic proteins
Transgenic technology in the production of therapeutic proteins Transgenic technology represents a new generation of biopharmaceutical production system to meet the medical needs of the new millennium.
More informationNext-Generation Facilities for Monoclonal Antibody Production
Advancing Development & Manufacturing Facilities Next-Generation Facilities for Monoclonal Antibody Production Niels Guldager ELECTRONICALLY REPRINTED FROM JULY 2009 The biopharmaceutical industry faces
More information50 g 650 L. *Average yields will vary depending upon a number of factors including type of phage, growth conditions used and developmental stage.
3430 Schmon Parkway Thorold, ON, Canada L2V 4Y6 Phone: 866-667-4362 (905) 227-8848 Fax: (905) 227-1061 Email: techsupport@norgenbiotek.com Phage DNA Isolation Kit Product # 46800, 46850 Product Insert
More informationExciting Trends in Bioprocessing
Exciting Trends in Bioprocessing Alfred Doig and Susan Dana Jones, Ph.D. April 20, 2015 BioProcess Technology Consultants, Inc. 12 Gill Street, Suite 5450 Woburn, MA 01801 Exciting Trends in Bioprocessing
More informationHiPer Ion Exchange Chromatography Teaching Kit
HiPer Ion Exchange Chromatography Teaching Kit Product Code: HTC001 Number of experiments that can be performed: 5 Duration of Experiment: Protocol: 5-6 hours Storage Instructions: The kit is stable for
More informationHis Mag Sepharose excel. Ni Sepharose excel. HisTrap excel. gelifesciences.com
Data file 29-168-49 AC His Mag Sepharose excel Ni Sepharose excel HisTrap excel Affinity chromatography His Mag Sepharose excel and Ni Sepharose excel (Fig 1) are immobilized metal ion affinity chromatography
More informationHost cell proteins (HCPs) are
B i op r o c e s s Technical Improved HCP Quantitation By Minimizing Antibody Cross- Reactivity to Target Proteins Xing Wang, Thomas Schomogy, Kristine Wells, and Ned M. Mozier Host cell proteins (HCPs)
More informationPrepacked chromatography columns for ÄKTA systems
GE Healthcare Life Sciences Prepacked chromatography columns for ÄKTA systems Selection guide imagination at work Different techniques Ion exchange chromatography (IEX) IEX separates proteins with differences
More informationGE Healthcare. Hydrophobic Interaction and Reversed Phase Chromatography. Principles and Methods
GE Healthcare Hydrophobic Interaction and Reversed Phase Chromatography Principles and Methods Handbooks from GE Healthcare Antibody Purification Handbook 18-1037-46 The Recombinant Protein Handbook Protein
More informationThe Importance of Developing a High Yield of Product
European Antibody Congress Lyon, 3 rd November 2005 The Importance of Developing a High Yield of Product John Birch, Lonza Biologics plc Monoclonal Antibodies A Success Story Fastest growing segment of
More informationAspects of industrial purification of peptides using large-scale chromatography. Lars Andersson and Jonas Persson
Aspects of industrial purification of peptides using large-scale chromatography Introduction By Lars Andersson and Jonas Persson PolyPeptide Laboratories (Sweden) AB PO Box 30089 SE-200 61 LIMHAMN SWEDEN
More informationReading instructions to partitioning in aqueous two-phase systems
Reading instructions to partitioning in aqueous two-phase systems Copy from Separation Processes in Biotechnology (ed. Asenjo) Aqueous two-phase separations, Albertsson, Johansson och Tjerneld OH-pictures
More informationChapter 3.2» Custom Monoclonal
198 3 3.2 Custom Monoclonal 199 Mouse monoclonal antibody development Chapter 3.2» Custom Monoclonal 200 In vitro monoclonals expression service 201 Mouse monoclonal antibody additional services 202 Magnetic
More informationLuca Romagnoli, Ph.D. Business Development Manager
Modelli innovativi di produzione per lo sviluppo di un processo altamente qualitativo di farmaci biologici Luca Romagnoli, Ph.D. Business Development Manager BIOLOGICAL DRUGS - SOURCES Monoclonal antibodies
More informationExpression and Purification of Recombinant Protein in bacteria and Yeast. Presented By: Puspa pandey, Mohit sachdeva & Ming yu
Expression and Purification of Recombinant Protein in bacteria and Yeast Presented By: Puspa pandey, Mohit sachdeva & Ming yu DNA Vectors Molecular carriers which carry fragments of DNA into host cell.
More informationViral Safety of Plasma-Derived Products
Viral Safety of Plasma-Derived Products SLIDE 1 This presentation will cover viral validation studies for plasma-derived products. FDA requires that the manufacturing process for biopharmaceutical products
More informationSingle Step Clarification & Capture of a Recombinant Protein from E. coli Osmotic Shock by Crossflow Chromatography
www.natrixseparations.com Complex Separations Single Step Clarification & Capture of a Recombinant Protein from E. coli Osmotic Shock by Crossflow Chromatography AN1000 Summary Recombinant Shiga toxin
More informationLFB GROUP & SANOFI combine their bioproduction capabilities to provide integrated CMO services for biopharmaceuticals
LFB GROUP & SANOFI combine their bioproduction capabilities to provide integrated CMO services for biopharmaceuticals MAbLaunch TM is a joint bioproduction platform combining LFB Biomanufacturing (LFB
More informationData File. Sephadex G-25 media and pre-packed columns. Introduction. Sephadex G-25 Bead structure. Desalting/buffer exchange and gel filtration
P H A R M A C I A B I O T E C H Sephadex G-25 media and pre-packed columns Data File Desalting/buffer exchange and gel filtration Reproducible desalting and buffer exchange in minutes with 90% 100% recovery
More informationGenomic DNA Extraction Kit INSTRUCTION MANUAL
Genomic DNA Extraction Kit INSTRUCTION MANUAL Table of Contents Introduction 3 Kit Components 3 Storage Conditions 4 Recommended Equipment and Reagents 4 Introduction to the Protocol 4 General Overview
More informationProtein Synthesis and Purification: Microbial Versus Mammalian Systems
STREAMLINING RECOMBINANT PROTEIN PRODUCTION The pharmaceutical industry is undergoing a deep transformation from small molecule drugs to biologics. Over the last decade, the percentage share of biologic-based
More informationTHOMAS C. RANSOHOFF Vice President and Principal Consultant
THOMAS C. RANSOHOFF Vice President and Principal Consultant +1.781.281.2704 (o) +1.617.759.4109 (m) transohoff@bptc.com Profile An expert in the development and scale up of biopharmaceutical processes;
More informationAccelerated Stability During Formulation Development of Early Stage Protein Therapeutics Pros and Cons of Contrasting Approaches
Accelerated Stability During Formulation Development of Early Stage Protein Therapeutics Pros and Cons of Contrasting Approaches 2008 IBC Formulation Strategies for Protein Therapeutics Tim Kelly, Ph.D.
More informationGE Healthcare Life Sciences
GE Life Sciences Controlled Order Intake Process for GE s Whatman Cellulose-Based Chromatography Media: Affected Products and Alternatives Product Code Item Description Discontinued or Transferred to Alternative
More informationAurum Ion Exchange Mini Kits and Columns. Instruction Manual
Aurum Ion Exchange Mini Kits and Columns Instruction Manual Catalog # 732-6710 Aurum AEX Mini Kits, 2 pk 732-6705 Aurum AEX Mini Kits, 10 pk 732-6706 Aurum AEX Mini Columns, 25 pk 732-6707 Aurum AEX Mini
More informationInterim Progress Report R&D Project 348. Development of a Field Test Kit for Detection of Blue-Green Algal Toxins
Interim Progress Report R&D Project 348 Development of a Field Test Kit for Detection of Blue-Green Algal Toxins Biocode Limited November 1992 R&D 348/04/A ENVIRONMENT AGENCY 135357 CONTENTS SUMMARY KEYWORDS
More informationIntegrated Protein Services
Integrated Protein Services Custom protein expression & purification Version DC04-0012 Expression strategy The first step in the recombinant protein generation process is to design an appropriate expression
More informationAccelerating drug development to FTIH: Potential of new expression technologies
Accelerating drug development to FTIH: Potential of new expression technologies Lekan Daramola Associate Director Biopharmaceutical Development, Cell Culture & Fermentation Sciences CMC Strategy Forum
More informationAssay Qualification Template for Host Cell Protein ELISA
Assay Qualification Template for Host Cell Protein ELISA Introduction: With ever increasing importance being placed on host cell protein (HCP) removal during the purification process, it is critical to
More informationAdvanced BioDesign Outlines Solutions. Antibody Overview. by Advanced BioDesign. Project Start. Immunogenicity. Selecting Your Antigen
Advanced BioDesign Outlines Solutions by Advanced BioDesign Antibody Overview Launching an immunisation programme is an important experimental step that needs care. With Advanced BioDesign, you may develop
More informationA World of Biomanufacturing: Shortages or Global Glut?
A World of Biomanufacturing: Shortages or Global Glut? Howard L. Levine, Ph.D. BioProcess Technology Consultants, Inc. BioProcess International Conference Vienna, Austria May 19-20, 2010 Steady growth
More informationApplication Note. Determination of Nitrite and Nitrate in Fruit Juices by UV Detection. Summary. Introduction. Experimental Sample Preparation
Application Note Determination of Nitrite and Nitrate in Fruit Juices by UV Detection Category Food Matrix Fruit Juice Method HPLC Keywords Ion pair chromatography, fruit juice, inorganic anions AZURA
More informationINSTRUCTION Probemaker
INSTRUCTION Probemaker Instructions for Duolink In Situ Probemaker PLUS (Art. no. 92009-0020) and Duolink In Situ Probemaker MINUS (Art. no. 92010-0020) Table of content 1. Introduction 4 2. Applications
More informationMarmara Üniversitesi Fen-Edebiyat Fakültesi Kimya Bölümü / Biyokimya Anabilim Dalı PURIFICATION AND CHARACTERIZATION OF PROTEINS
EXPERIMENT VI PURIFICATION AND CHARACTERIZATION OF PROTEINS I- Protein isolation and dialysis In order to investigate its structure and properties a protein must be obtained in pure form. Since proteins
More informationHisTrap HP, 1 ml and 5 ml
instructions HisTrap HP, 1 ml and 5 ml Caution! Contains nickel. May produce an allergic reaction. HiTrap affinity columns HisTrap HP is a prepacked, ready-to-use column for the preparative purification
More informationTrends in Upstream and Downstream Process Development for Antibody Manufacturing
Bioengineering 2014, 1, 188-212; doi:10.3390/bioengineering1040188 OPEN ACCESS bioengineering ISSN 2306-5354 www.mdpi.com/journal/bioengineering Review Trends in Upstream and Downstream Process Development
More informationReview of Chemical Equilibrium 7.51 September 1999. free [A] (µm)
Review of Chemical Equilibrium 7.51 September 1999 Equilibrium experiments study how the concentration of reaction products change as a function of reactant concentrations and/or reaction conditions. For
More informationOptimal Conditions for F(ab ) 2 Antibody Fragment Production from Mouse IgG2a
Optimal Conditions for F(ab ) 2 Antibody Fragment Production from Mouse IgG2a Ryan S. Stowers, 1 Jacqueline A. Callihan, 2 James D. Bryers 2 1 Department of Bioengineering, Clemson University, Clemson,
More informationThe Relationship between ph and Deionized Water
The Relationship between ph and Deionized Water The basics of ph The topic of ph and water has been well documented over the years; however, there is still much confusion about its significance in high
More informationEMABling Antibody Production Platform
EMABling Antibody Production Platform An industrial solution for the production of therapeutic antibodies with high cytotoxic activity B IOMANUFACTURING EMABling : A fully integrated development platform
More informationAnalyzing antibody sequence for recombinant antibody expression. Hangxing Yu, Ph.D Senior Scientist, GenScript May 20, 2015
Analyzing antibody sequence for recombinant antibody expression Hangxing Yu, Ph.D Senior Scientist, GenScript May 20, 2015 Presentation Outline 1 2 3 4 Antibody basics, structure and function Antibody
More informationFIDA for Rapid Detection of Protein Based Biomarkers
FIDA for Rapid Detection of Protein Based Biomarkers Guisheng Zhuang 1, Nicklas N. Poulsen 1, Nina Z. Andersen 1, Jesper Østergaard 1,2, Jørgen Schøller 2 and Henrik Jensen 1,2 1 Department of Pharmacy,
More informationYour partner in immunology
Your partner in immunology Expertise Expertise Reactivity Reactivity Quality Quality Advice Advice Who are we? Specialist of antibody engineering Covalab is a French biotechnology company, specialised
More informationRepligen Reports Third Quarter 2015 Financial Results
Repligen Corporation 41 Seyon Street Building #1, Suite 100 Waltham, Massachusetts 02453 Repligen Reports Third Quarter 2015 Financial Results - Product Sales Increase 31% to $19.8 Million - - Conference
More informationGeniron. Custom Antibody Services. Serum services Antibody Monoclonal. Purification Antibody Mono Y Genetic Immunization Genbody Polyclonal Antibody
Geniron Custom Antibody Services Serum services Antibody Monoclonal Purification Antibody Mono Y Genetic Immunization Genbody Polyclonal Antibody Geniron Poly Y WE PROVIDE OUR SERVICES TO With Expertise
More informationIntegrated Protein Services
Integrated Protein Services Custom protein expression & purification Last date of revision June 2015 Version DC04-0013 www.iba-lifesciences.com Expression strategy The first step in the recombinant protein
More informationMultiple Products in a Monoclonal Antibody S88.01 Batch Plant
Presented at the World Batch Forum North American Conference Chicago, IL May 16-19, 2004 900 Fox Valley Drive, Suite 204 Longwood, FL 32779-2552 +1.407.774.0207 Fax: +1.407.774.6751 E-mail: info@wbf.org
More informationMonoclonal Antibody Fragment Separation and Characterization Using Size Exclusion Chromatography Coupled with Mass Spectrometry
Monoclonal ntibody Fragment Separation and haracterization Using Size Exclusion hromatography oupled with Mass Spectrometry uthors Haiying hen Katherine McLaughlin Sepax Technologies, Inc. 5 Innovation
More informationAG 501-X8 and Bio-Rex MSZ 501(D) Mixed Bed Resin Instruction Manual. Bio-Rad Laboratories, 2000 Alfred Nobel Dr., Hercules CA 94547 LIT205 Rev B
AG 501-X8 and Bio-Rex MSZ 501(D) Mixed Bed Resin Instruction Manual Bio-Rad Laboratories, 2000 Alfred Nobel Dr., Hercules CA 94547 LIT205 Rev B Introduction Mixed bed resins are used for deionizing water
More informationINSTRUCTIONS 56-1190-98. Edition AC
Sephacryl S-100 High Resolution Sephacryl S-200 High Resolution Sephacryl S-300 High Resolution Sephacryl S-400 High Resolution Sephacryl S-500 High Resolution INSTRUCTIONS Sephacryl High Resolution chromatography
More informationHisTrap HP, 1 ml and 5 ml
instructions HisTrap HP, 1 ml and 5 ml Caution! Contains nickel. May produce an allergic reaction. HiTrap affinity columns HisTrap HP is a prepacked, ready-to-use column for the preparative purification
More informationSize Exclusion Chromatography
Size Exclusion Chromatography Size Exclusion Chromatography Instructors Stan Hitomi Coordinator Math & Science San Ramon Valley Unified School District Danville, CA Kirk Brown Lead Instructor, Edward Teller
More information