Continuous bioprocessing with Ultra-high productivity to expedite biologics development

Ultra-high productivity continuous bioprocesses have been developed for production of various biologics including monoclonal antibodies, fusion proteins and bispecfic antibodies. This enables, for example, 2,000L disposable bioreactors to achieve comparable productivity as traditional 20,000L stainless bioreactors, and to significantly reduce manufacturing cost of goods. This process technology platform consists of continuous cell culture and continuous direct product capture, and is being scaled up and implemented for production of clinical materials. Several case studies, which achieved a cell culture productivity of 2-3 g/L/day, and a similar purification yield of the traditional purification process, will highlight advantages of this integrated continuous process platform in terms of productivity gains and speed to clinic. Scale up and implementation challenges will also be discussed

Challenges and successes in developing and manufacturing multiple formats of bispecific antibodies

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Scale up and implementation of a high density long-term perfusion suspension cell culture in a 250L single use bioreactor

As part of efforts to develop a continuous processing platform for biologics manufacturing using a single use bioreactor, we have been focusing on development of several initial unit operations: high density perfusion suspension cell cultures and early product capture steps, in order to realize its potentials of being flexible, improving product quality and lowering costs. These initial unit operations require large volumes and represent the most important part of this processing platform. By integrating these steps into a continuous operation, we can deliver the largest benefits of this processing platform. In this presentation, we will discuss our efforts towards continuous biologics manufacturing using a case study. In particular, we focuses on direct scale-up of an ATF (Alternating Tangential Flow) based high density perfusion cell culture, from 2L scale coupled with ATF2 to 200L scale in a 250L single use bioreactor coupled with ATF6. Appropriate considerations of agitation and aeration rates, ATF operation parameters as well as bioreactor processing conditions resulted in successful scale-up of 100 folds. This high density long-term perfusion suspension cell culture was successfully implemented in a 250 single use bioreactor. Results of multiple completed batches indicate highly consistent process performance, productivity and quality of directly captured products. Furthermore, a representative scale-down model based on ATF2 was established for manufacturing support and further process optimization

Scale-up and scale-down challenges for a high density long-term perfusion suspension cell culture in large-scale single use bioreactors

As part of efforts to develop a continuous processing platform for biologics manufacturing using single use bioreactors, we have been focusing on development of several initial unit operations: high density perfusion suspension cell cultures and early product capture steps, in order to realize its potentials of being flexible, improving product quality and lowering costs. These initial unit operations require large volumes and represent the most important part of this processing platform. By integrating these steps into a continuous operation, we can deliver the largest benefits of this processing platform. In this presentation, we will discuss our efforts towards continuous biologics manufacturing using a case study focusing on direct scale-up of an ATF (Alternating Tangential Flow) based high density perfusion cell culture, from 2L scale coupled with ATF2 to large single use bioreactors coupled with ATF6 or ATF10. Appropriate considerations of agitation and aeration rates, ATF operation parameters as well as bioreactor processing conditions resulted in successful scale-up of more than 100 folds. This high density long-term perfusion suspension cell culture was successfully implemented in large scale single use bioreactors. Results of large-scale manufacturing batches indicate highly consistent process performance, productivity and quality of directly captured products. Furthermore, a representative scale-down model based on ATF2 was established for manufacturing support and further process optimization

Continuous bioprocessing for biologics manufacturing

Various strategies are explored for continuous bioprocessing for manufacturing of recombinant proteins and monoclonal antibodies: 1. continuous perfusion culture plus batch product capture; 2. continuous perfusion culture plus semi-continuous product capture; 3. continuous perfusion culture plus continuous product capture (Figure 1). Our main focus has been on the several initial unit operations: high cell concentration perfusion cell cultures and early product capture steps, in order to realize its potentials of being flexible, improving product quality and lowering costs. These initial unit operations require large volumes and represent the most important part of this processing platform, where we can deliver the largest benefits. Please click Additional Files below to see the full abstract

Rapid protein production using CHO cells: From transfection to 100g in 6 weeks

As more and more companies start to use pool materials for toxicology study to shorten overall IND enabling biologics development timeline, a rapid protein expression system using CHO-K1 cells has been developed to meet the demand for large amount of protein required for toxicology studies. 100-200g of proteins can be delivered 6 weeks after transfection using this CHO-K1 based expression system we call WuXian Express. The poster will describe this system and compare key antibody product quality attributes from materials generated from the WuXian Express, stable pools and Master Cell Banks (MCBs), and demonstrate comparability among them. Materials generated from this system have also been used at WuXi Biologics for developability study during antibody candidate molecule selection, early downstream, formulation and analytical method development, saving about 2 months compared with materials generated from stable pools

Challenges of scale down model for disposable bioreactors: Case studies on growth & product quality impacts

Despite wide-spread use of disposable bioreactors, there is a lack of well-established scale-down model for larger scale SUBs. Here we report a case of NS0 cell culture process transfer from 2000L stainless steel bioreactor (SST) to 2000L disposable bioreactor (SUB). Initial attempts in trying to grow the NS0 cells in the small scale 2D bags yielded non-satisfactory results, as growth was impacted by bag material type as well as by suppliers of the same bag material type. However, 3D bags of 50L and above proved to be supportive of the NS0 cell line growth. Even for cell lines that do not have growth issues in SUBs, surprising product quality difference between SUBs and traditional bench top glass bioreactors are still being observed, thus making the bench top glass bioreactors non-ideal as scale down models. We report two cases where glycan profiles of the expressed antibody products show such dramatic differences. In one case, extensive testing of glass bioreactors from various suppliers led to a particular type being able to mimic the glycan profiles from the SUB, whereas in the other case, alternative scale down model had to be identified and the process had to be modified to maintain the glycan profiles when scaling up to the 200L SUB

The effect of dopant and optical micro-cavity on the photoluminescence of Mn-doped ZnSe nanobelts

Pure and Mn-doped ZnSe nanobelts were synthesized by a convenient thermal evaporation method. Scanning electron microscopy, X-ray powder diffraction, energy dispersive X-ray spectroscopy and corresponding element mapping, and transmission electron microscope were used to examine the morphology, phase structure, crystallinity, composition, and growth direction of as-prepared nanobelts. Raman spectra were used to confirm the effective doping of Mn(2+) into ZnSe nanobelts. Micro-photoluminescence (PL) spectra were used to investigate the emission property of as-prepared samples. A dominant trapped-state emission band is observed in single ZnSe(Mn) nanobelt. However, we cannot observe the transition emission of Mn ion in this ZnSe(Mn) nanobelt, which confirm that Mn powder act as poor dopant. There are weak near-bandgap emission and strong (4)T(1) → (6)A(1) transition emission of Mn(2+) in single [Formula: see text] and [Formula: see text] nanobelt. More interesting, the (4)T(1) → (6)A(1) transition emission in [Formula: see text] nanobelt split into multi-bands. PL mapping of individual splitted sub-bands were carried out to explore the origin of multi-bands. These doped nanobelts with novel multi-bands emission can find application in frequency convertor and wavelength-tunable light emission devices

Further accelerating cell Line development and CMC timeline: The journey from COVID to non-COVID programs

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Ultra-intensified intermittent-perfusion fed-batch (UIIPFB) process quadrupled productivity of a bispecific antibody

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