MCC Applications

MCC Applications

List of MCC Applications 

ABSTRACT:

Here, we present a case study demonstrating the development of an MCC process for the Protein A affinity step of a monoclonal antibody (mAb) purification on the Octave BIO system and the straightforward scale-up to a clinical scale using the Octave PRO system. This was a joint study conducted between Catalent and Tosoh, where a direct comparison was made of a typical batch process with that of an MCC process while maintaining the same buffer conditions.

ABSTRACT:

Antibody fragments, such as antigen-binding domains (Fab), single chain variable fragments (ScFv) and heavy chain variable domains (nanobodies) have emerged as increasingly important therapeutic and diagnostic alternatives to full-length mAbs for a multitude of diseases. Protein L affinity chromatography is typically used for the capture of antibody fragments containing Fab part since they generally lack affinity to protein A ligand. With proteolytically produced Fab fragment as an example, here we compare two different protein L chromatography resins for the capture step in a batch mode. We also demonstrate how the transfer from a conventional batch mode to a multi-column continuous chromatography (MCC) process significantly increases the productivity of the Fab fragment capture. This leads to resin savings while simultaneously decreasing buffer consumption and consequently lower process operation costs. The complete bench-top protein L-based chromatography MCC platform presented here can feasibly be expanded to a pilot or manufacturing scale.

ABSTRACT:

Monoclonal antibody (mAb) drug biomanufacturers typically have relied on multi-step processes using batch-mode for optimal removal of impurities such as host-cell proteins (HCPs), DNA, adventitious viruses, and aggregates. However, additional purification steps increase downstream expenses significantly, including costs of supplementary resin, hardware, buffers, and area demand. Thus, it is imperative to design and test effective purification procedures for high-quality biotherapeutics, but with reasonable process costs, time, and manufacturing space requirements. Innovations in downstream mAb processing technology, such as the use of multi-column continuous chromatography (MCC) instrumentation, have recently been shown to significantly reduce operational costs, footprint, and time investment by increasing process productivity. To demonstrate the benefits of MCC technology in downstream processing, here we describe a two-step MCC platform for mAb purification using Tosoh Bioscience’s bench-top instrument, Octave® BIO, using SkillPak™ BIO pre-packed columns optimized for MCC applications.

ABSTRACT:

We describe a case study that directly compared a multi-column chromatography (MCC) process vs an optimized batch chromatography process for Protein A capture of a human monoclonal antibody at 100 g scale. The multi-column process was performed using a Semba ProPD™ Chromatography System with eight 50-ml columns (400 ml total resin volume). The batch process was performed with a GE AKTAprocess™ system with one 2.46 L column. Identical process solutions and protocols were used in each case. Each process was performed with two different Protein A resins; Toyopearl® AF rProtein A HC 650F and MabSelect SuRe™. Results demonstrated that the MCC process productivities were 3.3- and 3.1-fold higher with Toyopearl and MabSelect SuRe, respectively, than the corresponding batch processes. Buffer volumes used were 40% and 27% less, respectively, than the corresponding batch processes. When modeled at clinical scale (i.e. 2000 L) the MCC process represents savings of more than $800,000 in initial Protein A resin cost and 18 days per suite per year.

Catalent Webinar »

Comparing Continuous And Batch Processing In Downstream Purification »

ABSTRACT:

Multi-column continuous chromatography (MCC) offers significant economic advantages over traditional batch methods for purification of monoclonal antibodies (mAbs), including increased resin capacity utilization, smaller columns, reduced buffer consumption, and faster process time. The Protein A capture step is a primary target to apply MCC due to its high cost, which is driven even higher as improvements in upstream processing have produced a steady increase in mAb titers. In this study we consider the key factors in designing an MCC process that optimizes productivity. Process design begins with determination of the feed duration, which is governed by the mAb titer, column dimensions, residence time, binding capacity, and flow properties of the resin. Use of 3 columns in the capture zone enables efficient utilization of the resin even at short residence times, thereby increasing productivity. Column number and resin volume are optimized to relieve scheduling constraints experienced at high mAb titer while maximizing cycling for single use columns. Productivities exceeding 100 g mAb/L resin/h were achieved loading concentrated CHO expressed feed stream using a Semba ProPD™ System and 8-column MCC Protein A affinity process. Flow-through and bind-elute polishing steps including ion exchange and mixed mode have been converted from batch to MCC toward realization of a completely continuous biomanufacturing platform.

ABSTRACT:

We investigated a practical approach to optimizing a continuous Protein A capture process. Static and dynamic binding capacities were determined for various Protein A resins in single-column experiments followed by modeling the process in various column and flow configurations using a simple Excel-based tool. Models were tested using a lab-scale Octave® System. Residence time is the key driver of productivity and is subject to several constraints including flow velocity (pressure drop), mass transfer (dynamic capacity), and column number (process time). The highest productivities were obtained using 0.5 min residence time with 3 columns in the capture zone. For example, 97 g/L resin/h was achieved with high purity and recovery using TOYOPEARL^® AF rProtein A HC resin (Tosoh Bioscience). Optimal performance at low residence times requires adsorbents having high DBC and low pressure drop and MCC equipment that accommodates multiple process configurations. These considerations are important to maximize process economy at production scale at any feed titer.

ABSTRACT: 

We have used a lab-scale SMB/MCC instrument (Semba Octave® 10 System) to develop and optimize a continuous Protein A capture process. Here we present productivity data with five commercial Protein A adsorbents using feedstocks at 5 and 7.5 g/L mAb titers. With an 8-column process and appropriate capture resins, productivities approaching 100 g mAb/L resin/day were achieved with 7.5 g/L feedstock. In addition, we have examined hydroxyapatite and anion exchange as orthogonal MCC polishing steps for aggregate removal and concurrent depletion of impurities following Protein A capture. Results indicate that a completely continuous downstream process, including only two chromatographic steps, may be possible to further increase efficiency and reduce cost in mAb biomanufacture.