Antibody capture process based on magnetic beads from very high cell density suspension

Abstract

Cell clarification represents a major challenge for the intensification through very high cell density in the production of biopharmaceuticals such as monoclonal antibodies (mAbs). The present report proposes a solution to this challenge in a streamlined process where cell clarification and mAb capture are performed in a single step using magnetic beads coupled with protein A. Capture of mAb from non‐clarified CHO cell suspension showed promising results; however, it has not been demonstrated that it can handle the challenge of very high cell density as observed in intensified fed‐batch cultures. The performances of magnetic bead‐based mAb capture on non‐clarified cell suspension from intensified fed‐batch culture were studied. Capture from a culture at density larger than 100 × 106 cells/ml provided an adsorption efficiency of 99% and an overall yield of 93% with a logarithmic host cell protein (HCP) clearance of ≈2–3 and a resulting HCP concentration ≤≈5 ppm. These results show that direct capture from very high cell density cell suspension is possible without prior processing. This technology, which brings significant benefits in terms of operational cost reduction and performance improvements such as low HCP, can be a powerful tool alleviating the challenge of process intensification.

Proof-of-Concept of a Novel Cell Separation Technology Using Magnetic Agarose-Based Beads

“…The technology can also be applied to Protein L; however, this latter primarily binds to the κ-Fab fragment, so that it would potentially block the binding site of the cell-surface receptor and would therefore not be adequate for cell separation. Furthermore, the present study was a proof-of-concept exploiting our know-how of magnetic bead development and knowledge from a previous application for antibody purification process [22,23]. Potentially, other manufacturing processes, such as magnetic nanoparticles incorporated in hydrogel beads [29] could be used to manufacture such large magnetic particles.…”

“…Today, the relatively small volumes handled in IMS or FACS systems might be unsuitable to solve the bottleneck of large scale allogenic manufacturing [21]. In this context, previous studies using magnetic beads (MAGicBeads) of ≈100 µm radius, have provided an excellent performance to process large volumes of cell suspension and high cell concentrations [22,23].…”

“…A humanized monoclonal antibody IgG1, trastuzumab, recognizing the human epidermal growth factor receptor 2 (HER2/neu), was used for the separation of HER2 + cells. This antibody had been produced in house, in studies previously published [22,23].…”

Improving the reproducibility of size distribution of protein crystals produced in continuous slug flow crystallizer operated at short residence time

“…Anything but chromatography (ABC) methodologies, despite being argued to be changed to "Anything and Chromatography" (AAC) [78], has been widely investigated in the DSP of biopharmaceuticals [7,69]. Currently, membrane-based technology [77,[79][80][81], aqueous two-phase system (ATPS) [69,[82][83][84][85][86], magnetic separation [12,[87][88][89][90], precipitation [6,87,[91][92][93][94], and crystallization [6,26,38,84,[95][96][97][98] provide promising alternatives. Besides, to enhance the processing flexibility, better respond to fluctuating market demands, continuous manufacturing strategies have been sought as an effort to achieve process intensification in biologics manufacturing [11,70,[99][100][101][102].…”

Process intensification for CHO cell biomanufacturing

“…The clarification of its very high cell concentrations of up to 200×10 6 c•mL -1 combined with the purification of its very high product titres of up to 25 g•L -1 represent a major challenge. Recently, the application of ProtA coated magnetic beads for the purification of IgG from crude cell suspension has been reinvented 235 , applied in a cGMP-compliant set-up 236,237 and revealed promising results for this kind of intensified upstream process 238 and was patented 239 .…”