Developing an appropriate design space strategy to. With its focus on process development and largescale bioseparation tasks, this is tailormade reading for the professional bioengineer in both the biotech and pharmaceutical industries. Process development and scale up lfb biomanufacturing. The typical platform process is generally comprised of an initial capture step with protein a chromatography, a first polishing step with cationexchange media, and a final polishing step using anionexchange or multimodal media. As a consequence the bed height may be increased as long as the flowrate in units of columnvolumestime and load. Ngc mediumpressure chromatography systems with iqoq and cfrcompliant software chromatogtaphy columns and resins chromatogtaphy resin screening tools. Largescale monoclonal antibody purification by continuous. Improved efficiency and reproducibility in multidimensional multid purifications multid purification templates offer optimal efficiency for the tandem or sequential purification of multiple samples. Our protein production capabilities range from parallel screening of several constructs, through process development optimization, to scaleup manufacturing in e. Application note 28940349 aa highthroughput process. Their separation virtually always requires a gradient. By giorgio carta and alois jungbauer protein chromatography.
Rpc purification options and scale up with source media. Dec 20, 2019 a continuous integrated downstream process in which protein a chromatography capture, viral inactivation, flowthrough anion exchange, and mm cation exchange chromatography were integrated across two cadence biosmb pd multicolumn chromatography systems to purify a 25 l volume of harvested ccf. For the scaleup process, the rate of addition of granulation liquid is larger in amount for. The effect of the scaleup factors involved is studied. Process development, assay development, process optimization, scale up, cgmp manufacture commercia l manufactur e years. Scaleup of continuous multicolumn chromatography for the. The effect of the scale up factors involved is studied. Covers the basics of proteins properties, the unit operations involved in protein purification as well as important related issues of process design, integration with upstream operations, cost and. Divided into two large parts, protein chromatography.
Protein chromatography engineering fundamentals and measurement for process development and scaleup may 27th to june 1st 2018 an international chromatography course department of biotechnology university of natural resources and life sciences, vienna, austria registration form please fill in and return to. In the early stages of purification process development, different types of resins need to be tested at small scale 1. Except for the scaleup run using a pkl6 chromatography system from pall, we ran all tests on an akta explorer chromatography system from ge healthcare. Flexibility in scaleup can be achieved by employing the principle of scaling on a volume basis. The process is scaled by increasing the column diameter while keeping the bed height, the velocity, and the volumes of the different phases measured in column volumes constant. Chromatography columns and systems are critical factors to the successful separation of your valuable molecule. Those who downloaded this book also downloaded the following books. Development of purification processes for fully human. Process scaleup of protein and polypeptide separations. Process development and scale up, second edition presents all the necessary knowledge for effective process development in chromatographic bioseparation, both on small and large scale. For example, size exclusion chromatography sec is the key method used to quantify levels of protein aggregates and monomer. Process development and scale up on free shipping on qualified orders protein chromatography. Process development, assay development, process optimization, scaleup, cgmp manufacture commercia l manufactur e years. Lfb biomanufacturings services cover adaptation of cell line to serumproteinfree media and.
Integrating flexibility and scalability reliable flexible. Fast process development of a singlestep purification. Oct 01, 2011 we developed a highly efficient two step chromatography process by applying highthroughput process development htpd techniques using predictor plates for screening followed by optimization in smallcolumn format using a design of experiment doe approach figure 1. Scaleup studies in pharmaceutical products development chapter pdf available.
Multimodal chromatography handbook 29054808 ge healthcare life sciences nucleic acid sample preparation for downstream analyses principles and methods nucleic acid sample. Pdf scaleup studies in pharmaceutical products development. A comprehensive introduction to the art and science used by the biochemist and process engineer in the design and optimization of large scale protein purification processes. Both these methods are applied for the scale up of the developed lab scale whey protein resolution process. Protein chromatography process development and scaleup the authors giorgio carta university of virginia department of chemical engineering 102 engineers way charlottesville, va 229044741 usa alois jungbauer university of natural resources and applied life sciences boko department of biotechnology muthgasse 18 1190 vienna austria. Application guide for process development and scale p hydroxyapatite is formed from the chemical combination of calcium and phosphate salts. Sep 10, 2018 in this study, we focused on upscaling continuous protein a chromatography from lab to pilot scale using the cadence biosmb pd and the cadence biosmb process 80 system, respectively. The crystalline material so derived is widely used in analytical and preparative biomolecule research, as well as in industrial production of biologically active substances. Optimization and scaleup of hcicbased mab purification. Design of chromatographic processes giorgio carta university of virginia, department of chemical engineering, 102 engineers way, charlottesville, va 22904.
A practiceoriented allinone guide that provides the necessary knowledge for effective process development in chromatographic bioseparation, both for preparativescale and for largescale industrial applications. The biopharmaceutical discovery, development, and launch process. It should be noted that minicolumns do have several limitations as compared to labscale columns. Request pdf on oct 1, 2010, ruth freitag and others published protein chromatography.
An advantage in the area of mabs is the possibility of using a purification platform approach. The typical platform process is generally comprised of an initial capture step with protein achromatography, a first polishing step with cationexchange media, and a final polishing step using anionexchange or multimodal media. Developing an appropriate design space strategy to mitigate. Process development and scaleup, second edition presents all the necessary knowledge for effective process development in chromatographic bioseparation, both on small and large scale. Nov 05, 20 the scale up of a chromatographic separation is in principle a simple procedure since the process parameters are scalable in a linear fashion. The scaleup of a chromatographic separation is in principle a simple procedure since the process parameters are scalable in a linear fashion. Design of chromatographic processes protein chromatography. We provide standard and custom columns and systems from lab scale to pilot and process scale. A comprehensive introduction to the art and science used by the biochemist and process engineer in the design and optimization of largescale proteinpurification processes. Monoclonal antibody, monte carlo simulation, host cell protein, mabselect sure, capto adhere abstract we have developed and used a highly efficient monoclonal. Develop and finesse protein purification strategies across process development and scaleup using our extensive range of chromatography resins and instrumentation.
By giorgio carta and alois jungbauer freitag, ruth 20101001 00. The buffer blending valve automates optimal buffer formation to further accelerate your method development process. Alois jungbauer is the head of protein technology and downstream processing at the department of biotechnology of the university of natural resources and applied. These sites are distributed regularly throughout the crystal structure of the matrix.
Recent developments in bioprocessing of recombinant proteins. Optimization, robustness, and scaleup of mab purification. Scaleup of a downstream mab purification process using. Process scaleup of protein and polypeptide separations calculating the desorbing solvent or salt concentration adsorption of proteins and polypeptides on reversedphase columns is an allornothing phenomenon. Rpc purification options and scale up with source media appendix 1, extracted from hydrophobic interaction and reversed phase chromatography principles and methods pdf, ge healthcare, 2014 use source rpc media for purification and analysis of proteins, peptides and oligonucleotides. More detailed and sophisticated scaleup considerations are available in several books as well as in the recent chromatography literature. Ngc discover chromatography systems for method development. To determine purification feasibility of star substitutionbased bispecific antibodies, a scalable process for. Alois jungbauer is the head of protein technology and downstream processing at the department of biotechnology of the university of natural resources and.
Our protein production capabilities range from parallel screening of several constructs, through process development optimization, to scale up manufacturing in e. Chromatography systems and columns for every application. Additionally, we evaluated hardware and software capability whilst running the system for 10 days nonstop using feed from a perfusion bioreactor. Recent developments in bioprocessing of recombinant. Inspection of,, shows that the solution to is independent of the aspect ratio of the column provided the time scales t 0, t disp, t equil, and the adsorption isotherms q i c are identical. General chromatography column scale up approach bed height constant during scale up scale up by increasing column diameter and volumetric flow rate development scale to cgmp scale. Following a triedandtested concept, this guide has been developed over several years in training courses for biotech and chemical engineers in europe and the u. The downstream process starts with harvest clarification using a pod. He regularly organizes professional courses on various aspects of bioseparations, including a course on protein chromatography development and scale up together with alois jungbauer. Development of purification process for fully human bispecific antibodies based on isocratic positive mode affinity chromatography. The transfer of chromatographic processes from lab scale to pilot and production scale is an important and challenging task in the pharmaceutical. Covers the basics of proteins properties, the unit operations involved in protein purification as well as important related issues of process design, integration with upstream operations, cost and timing to market.
Development, modelling, optimisation and scaleup of. After completing this chapter, students will be able to. Fast process development of a singlestep purification using. Here we describe the optimization and robustness study as well as scaleup. By giorgio carta and alois jungbauer find, read and cite all the. Ceramic hydroxyapatite application guide for process. Using volumetric flow to scaleup chromatographic processes. Mar 24, 2018 those who downloaded this book also downloaded the following books. Lfb biomanufacturings services cover adaptation of cell line to serumproteinfree media and or media optimization for yield improvement. Effects of dispersion and adsorption kinetics on column performance in protein chromatography. Increasing capacity utilization in protein a chromatography. Both these methods are applied for the scaleup of the developed lab scale whey protein resolution process. At constant t 0 the linear flow velocity is proportional to the column length. Guidelines for optimization and scaleup in preparative.
For the scaleup process, the rate of addition of granulation liquid is. He regularly organizes professional courses on various aspects of bioseparations, including a course on protein chromatography development and scaleup together with alois jungbauer. A continuous integrated downstream process in which protein a chromatography capture, viral inactivation, flowthrough anion exchange, and mm cation exchange chromatography were integrated across two cadence biosmb pd multicolumn chromatography systems to purify a 25 l volume of harvested ccf. Following a tried and tested concept, this guide has been developed over several years in training courses for biotech and chemical engineers in europe and the u. High throughput process development htpd using prefilled predictor 96well filter plates for chromatography media and condition screening, followed by optimization of process conditions using akta avant 25 and unicorn 6, and then finetuning and scaleup using akta avant 150 system.
The objectives were a to implement a twostep continuous chromatography process supplemented by a final polishing step and b to design a platform for easy scale up and industrialization by employing only industrialcompatible conditions e. We have experience producing thousands of different proteins. When cgmp material for clinical trials and the market is produced, the lab scale purification process is scaled up based on methods acceptable to authorities in the various countries, including fda. Ngc chromatography systems for protein purification. Optimizing cationexchange chromatography with high. Application guide for process development and scaleup 3. Here we describe the optimization and robustness study as well as scale up. Agitation rate scaleup was based on constant power per unit volume of fluid pv, which translated to 100 rpm. We provide standard and custom columns and systems from labscale to pilot and processscale.