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. Fast process development of a singlestep purification using. The scaleup of a chromatographic separation is in principle a simple procedure since the process parameters are scalable in a linear fashion. 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. 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. Chromatography columns and systems are critical factors to the successful separation of your valuable molecule. In the early stages of purification process development, different types of resins need to be tested at small scale 1. Rpc purification options and scale up with source media. Scaleup of a downstream mab purification process using. More detailed and sophisticated scaleup considerations are available in several books as well as in the recent chromatography literature.
Recent developments in bioprocessing of recombinant. 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. 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. It should be noted that minicolumns do have several limitations as compared to labscale columns. By giorgio carta and alois jungbauer protein chromatography. Process development, assay development, process optimization, scaleup, cgmp manufacture commercia l manufactur e years. 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. 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. Application guide for process development and scaleup 3. 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. Their separation virtually always requires a gradient. Optimization and scaleup of hcicbased mab purification. Optimizing cationexchange chromatography with high.
Monoclonal antibody, monte carlo simulation, host cell protein, mabselect sure, capto adhere abstract we have developed and used a highly efficient monoclonal. Increasing capacity utilization in protein a chromatography. Fast process development of a singlestep purification. After completing this chapter, students will be able to. Those who downloaded this book also downloaded the following books. Scaleup of continuous multicolumn chromatography for the. For the scaleup process, the rate of addition of granulation liquid is larger in amount for. Integrating flexibility and scalability reliable flexible. We developed and scaled up a threecolumn purification process to meet the requirements for our 20l scale bioreactor run.
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. Flexibility in scaleup can be achieved by employing the principle of scaling on a volume basis. Pdf scaleup studies in pharmaceutical products development. We have experience producing thousands of different proteins. Largescale monoclonal antibody purification by continuous. 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.
Mar 24, 2018 those who downloaded this book also downloaded the following books. For example, size exclusion chromatography sec is the key method used to quantify levels of protein aggregates and monomer. The biopharmaceutical discovery, development, and launch process. He regularly organizes professional courses on various aspects of bioseparations, including a course on protein chromatography development and scaleup together with alois jungbauer. 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. Process development and scale up lfb biomanufacturing. Optimization, robustness, and scaleup of mab purification. A practiceoriented allinone guide that provides the necessary knowledge for effective process development in chromatographic bioseparation, both for preparativescale and for largescale industrial applications. Request pdf on oct 1, 2010, ruth freitag and others published protein chromatography.
As a consequence the bed height may be increased as long as the flowrate in units of columnvolumestime and load. 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. 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. With its focus on process development and large scale bioseparation tasks, this is tailormade reading for the professional bioengineer in both the biotech and pharmaceutical industries. The effect of the scale up factors involved is studied. Ceramic hydroxyapatite application guide for process. Chromatography systems and columns for every application. An advantage in the area of mabs is the possibility of using a purification platform approach. Improved efficiency and reproducibility in multidimensional multid purifications multid purification templates offer optimal efficiency for the tandem or sequential purification of multiple samples. The buffer blending valve automates optimal buffer formation to further accelerate your method development process. 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. A comprehensive introduction to the art and science used by the biochemist and process engineer in the design and optimization of largescale proteinpurification processes.
Guidelines for optimization and scaleup in preparative. Application note 28940349 aa highthroughput process. 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. 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. 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. Ngc mediumpressure chromatography systems with iqoq and cfrcompliant software chromatogtaphy columns and resins chromatogtaphy resin screening tools. At constant t 0 the linear flow velocity is proportional to the column length.
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. We provide standard and custom columns and systems from lab scale to pilot and process scale. Alois jungbauer is the head of protein technology and downstream processing at the department of biotechnology of the university of natural resources and applied. Development of purification processes for fully human. Process development and scale up on free shipping on qualified orders protein chromatography.
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. Lfb biomanufacturings services cover adaptation of cell line to serumproteinfree media and. Additionally, we evaluated hardware and software capability whilst running the system for 10 days nonstop using feed from a perfusion bioreactor. Process development and scaleup, second edition presents all the necessary knowledge for effective process development in chromatographic bioseparation, both on small and large scale. Design of chromatographic processes protein chromatography. Developing an appropriate design space strategy to mitigate. Scaleup studies in pharmaceutical products development chapter pdf available. Recent developments in bioprocessing of recombinant proteins. The transfer of chromatographic processes from lab scale to pilot and production scale is an important and challenging task in the pharmaceutical. 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. Ngc chromatography systems for protein purification. Multimodal chromatography handbook 29054808 ge healthcare life sciences nucleic acid sample preparation for downstream analyses principles and methods nucleic acid sample.
Rapid development and scaleup of biosimilar trastuzumab. The downstream process starts with harvest clarification using a pod. 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. Develop and finesse protein purification strategies across process development and scaleup using our extensive range of chromatography resins and instrumentation. Ngc discover chromatography systems for method development. The effect of the scaleup factors involved is studied. These sites are distributed regularly throughout the crystal structure of the matrix. Development, modelling, optimisation and scaleup of. Effects of dispersion and adsorption kinetics on column performance in protein chromatography. Both these methods are applied for the scale up of the developed lab scale whey protein resolution process. Agitation rate scaleup was based on constant power per unit volume of fluid pv, which translated to 100 rpm. Developing an appropriate design space strategy to.
Lfb biomanufacturings services cover adaptation of cell line to serumproteinfree media and or media optimization for yield improvement. 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. Here we describe the optimization and robustness study as well as scaleup. Development of purification process for fully human bispecific antibodies based on isocratic positive mode affinity chromatography. Using volumetric flow to scaleup chromatographic processes.
He regularly organizes professional courses on various aspects of bioseparations, including a course on protein chromatography development and scale up together with alois jungbauer. 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. Application guide for process development and scale p hydroxyapatite is formed from the chemical combination of calcium and phosphate salts. Process development, assay development, process optimization, scale up, cgmp manufacture commercia l manufactur e years. Both these methods are applied for the scaleup of the developed lab scale whey protein resolution process. The crystalline material so derived is widely used in analytical and preparative biomolecule research, as well as in industrial production of biologically active substances. To determine purification feasibility of star substitutionbased bispecific antibodies, a scalable process for.
Process scaleup of protein and polypeptide separations. 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. Design of chromatographic processes giorgio carta university of virginia, department of chemical engineering, 102 engineers way, charlottesville, va 22904. By giorgio carta and alois jungbauer find, read and cite all the. Process development and scaleup are essential to obtain a costeffective commercial product.
We provide standard and custom columns and systems from labscale to pilot and processscale. Divided into two large parts, protein chromatography. 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. The key parameter when scalingup is the mobile phase residence time t 0. Here we describe the optimization and robustness study as well as scale up. 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. Our protein production capabilities range from parallel screening of several constructs, through process development optimization, to scaleup manufacturing in e. 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. Our protein production capabilities range from parallel screening of several constructs, through process development optimization, to scale up manufacturing in e. Alois jungbauer is the head of protein technology and downstream processing at the department of biotechnology of the university of natural resources and. By giorgio carta and alois jungbauer freitag, ruth 20101001 00.