Table 1: Virus hemagglutinin (HA) yield, TCID 50, DNA, total protein, and host cell protein (HCP)/HA quotients in a purification scheme incorporating microfiltration and two-step chromatography using Capto DeVirS and Capto Core 700 The resulting total cost per batch for this step could be lowered by almost two-fold. As a consequence, labor costs, buffer costs, and costs for Capto Core 700 medium were significantly lower than for traditional gel filtration. The higher load volumes achieved with Capto Core 700, relative to gel filtration, resulted in smaller volumes of chromatography medium to process the same amount of material within a shorter time-frame. (Further examples of purification with Capto Core 700 can be found in reference 2).Ĭost Analysis of Capto Core 700 versus gel filtrationīased on the running conditions and results for the Capto Core 700 example in Table 1, and from experience of using gel filtration, a cost comparison for processing a feed from a 1000 Lfermentation is presented in Table 2. The infectivity of the virus was retained throughout the process, as shown by the titer measured with TCID 50.
Capto Core 700 further reduced protein levels by three- to five-fold. DNA was reduced 2.8 log and proteins five- to seven-fold by Capto DeVirS. In this case, good yield of virus HA as well as significant removal of HCP and DNA were observed. Table 1 shows the results in terms of hemagglutinin (HA) recovery, TCID 50, DNA, and protein removal at each step of the process. This reduces the need for optimization such as buffer exchange or dilution between steps (even with different feed materials). The robust performance of the multimodal octylamine ligand in a relatively wide range of NaCl concentrations and pH (Figure 1) gives Capto Core 700 a wide window of operation. Protein Binding Capacity and the Operating Window In addition, the impact of Capto Core 700 on process economy was compared to that of Sepharose™ 4 Fast Flow, a gel filtration medium often used in the vaccine manufacturing industry. An experiment to determine the window of operations for influenza virus purification was performed using PreDictor™ 96-well plates for high-throughput process development. This study evaluates Capto Core 700 as an alternative to current chromatography technologies used in vaccine processes. These ligands bind various contaminants strongly over a wide range of pH and salt concentrations. The octylamine ligands in the core of the bead are multimodal, being both hydrophobic and positively charged. Viruses and other large entities with a molecular mass (M r) greater than approximately 700 000 (700 kDa) are excluded and are collected in the flowthrough. Small contaminant molecules enter into the beads where they are captured. The Capto Core 700 medium has a core bead design and consists of an inactive outer layer and a ligand-activated core. Clearly, new techniques are needed to improve productivity and economy in vaccine production.Ĭapto™ Core 700 is a member of GE Healthcare Life Sciences’ Capto line of chromatography media ( 1). In these processes, both sucrose density gradient ultracentrifugation and gel filtration have limitations in scalability and throughput. The methods for purification processes have typically involved a combination of sucrose density gradient ultracentrifugation, ultrafiltration/diafiltration (UF/DF) with hollow-fiber membranes, and chromatography using affinity, ion-exchange, and/or gel filtration media (resins). However, to meet the needs for pandemic preparedness and the scalability of vaccine production, cell-based processes are now being developed and implemented in the industry. Influenza vaccine has historically been produced in embryonated chicken eggs.
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