Biopharmaceutical industry is increasingly moving towards use of disposable single use systems (SUS) for development as well as manufacture of a wide range of vaccines, therapeutic proteins and mAbs. Biopharmaceutical processes involve multiple steps with a multitude of process intermediates with different process conditions and objectives at each step.
Single Use Systems (SUS) offer multiple advantages of reduced capital expenditure, reduced change over time and increased process flexibility while doing away with expensive and time consuming CIP/SIP procedures and validation requirements associated with reusable stainless steel systems.Aseptically connecting single use components to form a usable assembly or to connect a SUS to another single use or a reusable system, for example a sterile media bag to a bioreactor, is a censorious requirement.
Gamma Sterile AseptiLink SC, genderless connectors are designed to provide a fast and convenient aseptic connection between two processing streams, such as a container to a sampling line, media bags to a bioreactor or a filtration assembly to a filling line, without the need of a biocontainment hood. In other words, it provides a validated robust connecting mechanism within sterile as well as non sterile environments, while eliminating the need for sophisticated and complicated hardware.
Bio-pharmaceuticals R&Ds are working with mammalian cell expression systems for developing mAbs and other therapeutic proteins. Scientists today are using increasingly smaller culture volumes grown in 10ml to 15ml micro bioreactor formats or even smaller volumes in 24 to 48 plate systems for clone and media optimization, cell line selection, small scale perfusion mimic and early process optimization. In these applications multiple cell harvests need to be clarified to remove cells and debris before the critical metabolite analysis/ chromatography steps. Conventional methods for clarification of such small volumes (few mL) are centrifugation followed by 0.2µm membrane filtration.
However centrifugation is a time consuming, cumbersome process and may also result in shearing of cells resulting in increased cell debris, cell organelles and intracellular proteins in the supernatant. The conventional syringe filter, although used for final filtration of the supernatant does not offer an efficient solution in terms of throughputs.’
Closed loop sterility testing systems have been successful in prevention of false positives while sterility testing of sterile injectables. However, there are special containers such as insulin cartridges (that fit into hand held delivery systems), wherein increased manual intervention is required for pooling the same into a 100ml vial before these can be transferred/filtered through the closed sterility test canisters. This increases the possibility of extraneous contamination and thereby false positives.
A syringe is used to draw the insulin sample from the specialized cartridge and is then pooled into a 100ml vial to be transferred/filtered through the sterility test canisters.
This involves multiple risks and cons such as:
Chances of extraneous contamination, spillage and manual error
Clarification of turbid solutions is a key requirement to achieve critical objectives in the pharmaceutical and bio-pharmaceutical manufacturing which range from clarification, polishing, bio-burden reduction to sterilization of process fluids.
These process streams can range from easy to filter, predictable solutions such as large volume parenteral, water for injection and buffers, to difficult to filter colloidal solutions, emulsions, liposomal drug delivery systems, cell cultures, lysates, plasma, etc.
0.2 µm Membrane filters are used at various stages of the manufacturing process and fluids with a wide ranging contamination profile, including difficult to filter colloids and compressible particles, pose a serious challenge to these filters. This leads to lower throughputs and higher filtration footprint.
Biopharmaceutical manufacturing processes involve multiple process steps with a wide variety of process streams, including some with the most challenging contamination profiles. These contaminants range from cell fragments, cell organelles, colloids and lipids to very fine protein precipitates. Some of these process steps use filter aids which although aid in retention of some of these contaminants, contribute their own fines into the downstream. As expensive and sophisticated equipment and consumables are used for downstream purification, such high contamination profile process streams tend to rapidly clog the 0.2µm filters, normally used to protect these.
MDI has created an innovative new product called Aseptivac. We all have used vacuum filtration units to quickly filter a liter of buffer in the lab – but what happens when you have a difficult to filter solution, or the media is plugging the filter?
Bio-pharmaceutical manufacturing is a complex, multistep process involving a very wide variety of process streams under different process conditions at different steps. These process streams include cell culture media, media additives, growth regulators in the upstream and post centrifuge cell harvest supernatants, post viral inactivation process intermediates, buffers, and high value product concentrates in the downstream. Filtration and purification of such a wide spectrum of process streams, to achieve different process objectives at each process step, is quite a challenge for the process owner. Also the increasingly competitive market conditions, due to the introduction of biosimilars, further escalates the demand for higher standards of filter performance, with respect to economy, safety, and impact on final product quality.
Lab scale clarification of cell harvests is increasingly becoming more challenging as the standard clarification techniques such as centrifugation, tangential flow filtration (TFF) and depth filtration are not easily adaptable to micro bioreactors which require multiple processing of small volumes of cell harvests, typically 50 mL. Centrifugation may damage the shear sensitive mammalian cells and not only release undesirable intra cellular proteins and DNA into the broth but also submicron particles which are difficult to remove. TFF systems require long setup time and are prohibitively expensive for lab scale volumes. Depth filters on the other hand necessitate monitoring of differential pressures and sometimes allow passage of cell debris and other fine particulate which tend to choke the downstream 0.2µm sterilizing grade filters. Also, these require large pre-use flush volumes to remove inherent extractables owing to their cellulosic matrix with inorganic filter aids such as diatomaceous earth or perlite. Continue reading “Clarification of Cell Harvests from Micro Bioreactors”
Bio-pharmaceutical industry is involved in development and manufacture of therapeutic drug proteins and monoclonal antibiotics using mammalian cell expression systems, a vital and constantly developing technology.
Cultured cells from mammalian cell expression systems, ranging from a few mL to a few liters in shaker flasks, cell factories and small bioreactors to thousands of liters in large bioreactors, deliver extracellular protein and need to be harvested and clarified for downstream processing to obtain the purified protein of interest. Cell culture clarification is necessary to protect highly sensitive and expensive downstream processes such as diafiltration, ultrafiltration and protein chromatography, and is a challenging task as it contains whole cells, cell debris from dead cells as well a whole gamut of proteins from the cellular activity.