Integrated continuous biomanufacturing platform with ATF perfusion and one column chromatography operation for optimum resin utilization and productivity.

A new integrated continuous biomanufacturing platform for continuous production of antibodies at fixed cell volumes and cell concentrations for extended periods with immediate capture is presented. Upstream antibody production has reached technological maturity, however, the bottleneck for continuous biomanufacturing remains the efficient and cost-effective capture of therapeutic antibodies in an initial chromatography step. In this study, the first successful attempt at using one-column continuous chromatography (OCC) for the continuous capture of therapeutic antibodies produced through alternating tangential flow perfusion is presented. By performing upstream media optimizations, the upstream perfusion rate was reduced to one vessel volume per day (vv/d), increasing antibody titer and reducing the volume of perfusate. In addition, process improvements were performed to increase productivity by 80% over previously reported values. In addition, a real-time method for evaluating column performance to make column switching decisions was developed. This improved productivity coupled with the use of a single-column improved process monitoring and control in OCC compared to multi-column systems. This approach is the first report on using a single column for the implementation of an integrated continuous biomanufacturing platform and offers a cost-effective and flexible platform process for the manufacture of therapeutic proteins.

The Current Scientific and Regulatory Landscape in Advancing Integrated Continuous Biopharmaceutical Manufacturing.

There is a trend across the pharmaceutical sector toward process intensification and continuous manufacturing to produce small-molecule drugs or biotechnology products. For biotechnology products, advancing the manufacturing technology behind upstream and downstream processes has the potential to reduce product shortages and variability, allow for production flexibility, simplify scale-up procedures, improve product quality, reduce facility footprints, increase productivity, and reduce production costs. On the upstream side of biotechnology manufacturing, continuous perfusion cell cultures are fairly well established. However, truly integrated continuous biomanufacturing requires the uninterrupted connection of continuous unit operations (upstream and downstream) with no isolated intermediate or hold steps occurring between them. This work examines the current scientific and regulatory landscape surrounding the implementation of integrated continuous biomanufacturing.

Quantification of protein mixture in chromatographic separation using multi-wavelength UV spectra.

In therapeutic protein production, the protein purification with chromatographic processes is of high importance in separating the qualified proteins from the impurities for consistent product quality. Therefore, to aid real-time monitoring of the protein purification processes, various kinds of methodologies have been proposed until now. However, the majority of them still rely on the use of a single ultraviolet (UV) absorbance or the utilization of expensive and time-consuming instruments, thus requiring a simple, fast, and cost-effective methodology for protein quantification. In this study, the feasibility of using multiwavelength UV spectroscopy was investigated as an alternative tool for the real-time monitoring of the protein mixtures in protein purification. To this end, three different proteins were selected as a model system for the protein mixture, and the multivariate UV spectra were analyzed to construct the reliable quantification models for different proteins of interest. By using various chemometrics tools, such as partial least squares (PLS), the validity of estimating the protein concentration from the UV spectra of the mixture samples was rigorously analyzed with their prediction performance, and the results indicated that the multiwavelength UV spectra had sufficient sensitivity and accuracy to estimate the protein concentrations in mixture, demonstrating its usefulness for the rapid quantification of the protein mixtures in protein purification.