Protein A chromatography is a near-ubiquitous method of mAb capture in bioprocesses. The use low pH buffer for elution from protein known to contribute product aggregation. Yet, more limited set evidence suggests that may not be the sole cause aggregation chromatography, rather, other facets process significantly. This paper presents well-defined investigating this problem. An IgG4 was incubated after (typical viral inactivation hold) and quantity monomer neutralised samples determined by...

The characterization of host cell protein (HCP) content during the production therapeutic recombinant proteins is an important aspect in drug development process. Despite this, key components HCP profile and how this changes with processing has not been fully investigated. Here we have investigated supernatant at different times throughout culture a null model GS-CHO monoclonal antibody producing mammalian line grown fed-batch mode. Using 2D-PAGE LC-MS/MS identify number intracellular (e.g.,...

Fabricating nanofibres with reproducible characteristics is an important demand in the membrane industry order to establish commercial viability. In this study, effect of controlled atmospheric conditions on electrospun cellulose acetate (CA) was evaluated for temperatures ranging 17.5–32.5 °C and relative humidity 20–70%. CA solution (0.2 g/mL) a solvent mixture acetone/dimethylformamide/ethanol (2:2:1) into nonwoven fibre mesh diameter from 150 nm 1 μm. The resulting were analysed by...

Abstract Recombinant protein products such as monoclonal antibodies (mAbs) for use in the clinic must be clear of host cell impurities (HCP), DNA/RNA, and high molecular weight immunogenic aggregates. Despite need to remove monitor HCPs, nature, fate these during downstream processing (DSP) remains poorly characterized. We have applied a proteomic approach investigate dynamics HCPs supernatant mAb producing line early DSP including centrifugation, depth filtration, A capture chromatography....

Protein A chromatography is a critical and 'gold-standard' step in the purification of monoclonal antibody (mAb) products. Its ability to remove >98% impurities single alleviates burden on subsequent process steps facilitates implementation platform processes, with minimal number chromatographic steps. Here, we have evaluated four commercially available protein matrices terms their host cell proteins (HCPs), complex group related that must be removed levels. SELDI-TOF MS was used as...

The purpose of this study was to assess the stability protein formulations using a device designed generate defined, quantifiable levels shear in presence solid-liquid interface. device, based on rotating disk, produced strain rates up 3.4 × 104 s−1 (at 250 rps) and exclude air-liquid interfaces enable temperature be controlled. Computational fluid dynamics (CFD) used flow patterns within determine rate (s−1) at range disk speeds. then effect monoclonal IgG4 high Monomeric antibody...

An integrated experimental and modeling approach for the design of high productivity protein A chromatography is presented to maximize in bioproduct manufacture. The consists four steps: (1) small-scale experimentation, (2) model parameter estimation, (3) optimization (4) validation with process verification. use experimentation enables fewer experiments be performed, thus minimizes time materials required order gain understanding, which key importance during development. application...

Protein separation is an integral step in biopharmaceutical manufacture with diffusion-limited packed bed chromatography remaining the default choice for industry. Rapid bind-elute using convective mass transfer media offers advantages productivity by operating at high flowrates. Electrospun nanofibre adsorbents are a non-woven fibre matrix of surface area and porosity previously investigated as bioseparation medium. The effects compression layers, subsequent heat treatment after...

An ever increasing focus is being placed on the manufacturing costs of biotherapeutics. The drive towards continuous processing offers one opportunity to address these through advantages it offers. Continuous operation presents opportunities for real-time process monitoring and automated control with potential benefits including predictable product specification, reduced labour costs, integration other processes. Specifically chromatographic operations an use expensive media more efficiently...

A model based approach has been developed and used to identify robust operating conditions for an industrial hydrophobic interaction chromatography where resin lot variability, combined with feed stream was resulting in serious performance issues during the purification of a multi component therapeutic protein from crude material. An equilibrium dispersive formulated which successfully predicted key product critical quality attribute validation studies. The then parameter ranges that assured...

Virus-like particles (VLPs) are supramolecular protein assemblies with the potential for unique and exciting applications in synthetic biology medicine. Despite attention VLPs have gained thus far, considerable limitations still persist their production. Poorly scalable manufacturing technologies inconsistent product architectures continue to restrict full of VLPs. Cell-free synthesis (CFPS) offers an alternative approach VLP production has already proven be successful, albeit using extracts...

The majority of CD19-directed CAR T cell products are manufactured using an autologous process. Although a patient's leukapheresis reduces the risks rejection, it introduces variability in starting material composition and presence populations that might negatively affect production chimeric antigen receptor (CAR) cells, such as myeloid cells. In this work, effect monocytes (CD14) on level activation, growth, transduction efficiency was monitored across well plate culture bag platforms...

The purification of extracellular vesicles (EVs) remains a major hurdle in the progression fundamental research and commercial application EV-based products. In this study, we evaluated potential heparin affinity chromatography (HAC) to purify neural stem cell-derived EVs as part multistep process. Bind-elute chromatography, such HAC, is an attractive method because it highly scalable, robust can be automated. Our findings support interaction between heparin. recovery using HAC based on...

Traditional poly(ether sulfone) (PES) filters, widely used for sterile, viral, and ultrafiltration, often exhibit restrictions in their selectivity-permeability profile due to heterogeneous pore size distribution. This limitation has sparked interest developing novel isoporous membrane materials fabrication techniques. Among promising candidates, block copolymer (BCP) membranes produced via self-assembly nonsolvent-induced phase separation (SNIPS) offer significant advantages, including...