Figure 1:
Millipore Corporation’s new Viresolve Pro and Pro+ solutions are a major leap forward in virus clearance technology. As biotech manufacturers experience high titers, the evolution of process platforms and increasing focus on cost of goods, the demand for a robust high-capacity unit operation is increasing. Viresolve Pro and Pro+ solutions deliver robust parvovirus clearance, high productivity, and performance assurance.
The Viresolve Pro and Pro+ solutions offer the highest level of virus clearance for the development of biopharmaceutical products. The patent pending membranes, newly designed device formats, new highly sensitive performance tests, and comprehensive range of services and virus clearance expertise offer high throughput and virus clearance assurance. For challenging feedstreams, the Viresolve Pro+ solution includes an adsorptive polishing step that protects against variability in solution conditions. All device formats are fully disposable and caustic stable for optional in situ sa...
Figure 1:
Millipore Corporation’s new ChromaSorb membrane adsorber uses an innovative membrane-based anion exchanger to perform robust impurity clearance at a wide range of salt concentrations. This feature enables users to improve their process economics by eliminating the downstream bottleneck of buffer tank capacity inherent with the use of other membrane adsorbers, as well as traditional resin chromatography.
ChromaSorb membrane adsorber provides enhanced process flexibility and efficiency and a smaller manufacturing footprint than traditional anion resins and columns, eliminating the need to dilute feedstreams prior to processing. The ChromaSorb solution comprises a linearly scalable family of single-use devices, which bind negative impurities, including host cell protein (HCP), DNA, endotoxins, and viruses at salt concentrations well above those of traditional resins and membranes.
The ChromaSorb 0.08-mL device is the smallest chromatographic scaling device available, designed to minimize the use of...
Figure 1:
SterilEnz®-II/G connectors and SterilEnz Samplers represent a novel design improvement when compared with common fittings and fluid injection sites. This article briefly discusses the features, benefits, and technical aspects surrounding the design and integrity testing of the SterilEnz product line as well as advantages over their predecessors.
SterilEnz-II/G Flange Connectors and Samplers
Each SterilEnz-II/G connector is hermetically sealed inside a rugged overwrap pouch (Figure 1). The connector is fitted with a pre-attached, medical grade silicone gasket. Because of these unique features, single-use bags manufactured with SterilEnz-II/G connectors provide greater sterility assurance and ease of use.
Figure 1: ()
SterilEnz Samplers solve the common problem that occurs when using injection sites on single-use systems — that is, how does one maintain the sterility of the injection site itself in a nonsterile environment? The solution again is to seal the injection site inside a hermetic closure...
Figure 1:
Full-length antibody biopharmaceuticals continue to capture significant market due to their validated effectiveness as specific therapeutics. However, more recently it has been recognized that full-length antibodies may not always be necessary because antibody fragments can provide opportunity for new therapeutics with less complicated, higher-yielding production processes such as
Escherichia coli
and yeast microbial systems. For full-length antibodies, the downstream processing is facilitated by use of protein A–based affinity adsorbents for capture and initial purification; however there has been a lack of a similar specific adsorbent for the capture and initial purification of antibody fragments.
A synthetic ligand capable of binding to the light-chain binding site of antibodies was identified by screening of chemical combinatorial ligand libraries. The resulting adsorbent, Fabsorbent™ F1P HF, is effective for capture and initial purification of antibody fragments from a variety of sources.
Figure 1:
Dead-end filters using microporous membranes of synthetic polymers such as PES, polyamide, cellulose acetate, and PVDF are extensively used for microbial control in a wide range of biopharmaceutical liquid filtration applications. Typical applications include sterile media addition into bioreactors, cell harvest clarification, chromatography column protection, and final sterile filtration of purified bulk drug substances. Downstream processing in the biopharmaceutical industry requires multiple bioburden reduction steps that commonly use 0.2-µm sterilizing grade filters.
There are innovative application-specific approaches in sterilizing-grade membrane filtration such as Sartopore® 2 XLG 0.8/0.2 and Sartopore® 2 XLI 0.35/0.2 PES filters from Sartorius Stedim Biotech. Both filters have different prefilter membrane retention ratings designed for optimum performance with fluids having specific particle size distribution characteristics. These filters combine specialized prefilters with optimized pl...
Figure 1:
Single-use purification processes typically use peristaltic pumps to provide single-use fluid pathways. However, peristaltic pumps are known for generating severe pressure oscillations. These oscillations have an effect on the accurate measurement of the process, thus limiting the effective monitoring and controlling of system pressure.
SciLog is pleased to announce a significant performance breakthrough in pressure monitoring: achievement of a 98% reduction in signal noise level associated with peristaltic pump pulsation. In this solution, a significant increase in the signal to noise ratio is achieved while a fast sensor response is maintained. This important breakthrough enables automation applications and accurate data collection in single-use purification platforms.
SciLog manufactures pressure sensors for single-use purification applications in the bioprocess industry including tangential flow filtration (TFF), normal flow filtration (NFF), and preparative chromatography. SciLog pressure s...
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During the past few years the biotech industry has started to move away from protein A-based platform systems for MAb purifications. The high cost of protein A resins, their modest dynamic binding capacities (DBC), and their limited base stability place restrictions on a developmental engineer’s ability to reduce process costs and improve throughput for a given plant footprint.
Recent literature reports from Percivia (
1
) and Medarex (
2
) have described the use of improved cation exchange resins for MAb capture steps rather than more traditional protein A media. Scientists at Percivia were able to triple the loading capacity of the capture step with significant reduction in associated process costs, while process developers at Medarex, who incorporated precipitation with low pH conditioning for the feedstock, also reported loading capacities of up to 100 mg/mL resin (
2
).
These new cation exchange resins create a significant economic improvement parameter for developmental engineers.
Percivia...
Orbital welding has long been the most popular and accepted method for joining 316L stainless steel tubing within the bioprocessing and pharmaceutical sectors. Yet as requirements for orbital welding continue to expand, so does the need to select a system that will pay dividends in more ways than one. Recent advancements in small, portable inverter power supplies help improve the consistency and reliability of welds as well as operator efficiencies.
Open Platform Design
To enable optimal efficiencies, a power supply must be designed with an open platform that allows an interface with standard devices and the ability to expand as technology evolves. Today’s power supplies control aspects of the welding process that historically would be undertaken manually, such as travel speed, arc gap, and gas flow. This automation minimizes many variables that can lead to errors or defects and enables welders to focus on overseeing the process and completing welds. The result is a more efficient and streamlined process ...
Advanced Scientifics is a leader in the design and manufacturing of single-use systems for the pharmaceutical and biotechnology industries. The company offers a multitude of solutions for flexible containers. Numerous films are available such as polyethylene (PE) and ethyl vinyl acetate (EVA) bags, as well as C-Flex, Silicone, PVC, and PharMed BPT tubing types. With sizes ranging from 50 mL to 2,500 L, Advanced Scientifics can produce single-use containers of nearly any configuration as well as multiple porting options with 700+ components in-house to choose from. Advanced Scientifics also has the capability to produce 3-D bags from 20 L and up, specific to customer specifications and tote dimensions.
Safest Downstream Tubing Sets
Advanced Scientifics offers a wide assortment of transfer and filtration sets for most any application. Our sets are used throughout the cell culture process, from media or buffer transfer to seeding or harvest — to separation and purification — to final packaging. Advanced Scie...
Applikon Biotechnology is the world’s leading manufacturer of glass autoclavable vessels. Over 25 years ago, autoclavable vessels were sold as systems by many companies. When a large west-coast biotech company wanted a modular approach and also wanted changes in the design of the vessel, Applikon listened and complied. The glass bioreactor is now the standard of the industry and is sold in similar forms by many of Applikon’s competitors.
Applikon has kept its focus on the bioreactor and anything related to or working with the bioreactor. In doing so, Applikon has become the innovator in this segment of the market. Through listening and observing, Applikon has introduced the following innovative products to the market:
An Adaptive Business Philosophy
Applikon’s business philosophy has evolved to be open to change and willing to accept new technologies from sources outside the industry.
Historically, the bioreactor supplier market has had high rates of turnover. This is due to the complexity of the equipmen...
Figure 1:
ATMI® LifeSciences offers five different disposable mixing technologies that provide varied cost, operational, and performance advantages over conventional stainless steel mixing vessels and competing disposable mixers. Criteria for selecting the best ATMI mixing technology include scale, particle, and sheer sensitivity, mixing power, physical fit, and economic considerations. This application note illustrates how the LevTech® Magnetic Mixing system performs in a demanding media mixing application.
Introduction
The LevTech Magnetic Mixer is a compact and noninvasive single-use mixing system. The mixing bag incorporates a bottom-mounted, magnetically driven impeller that provides high-torque mixing for powder–liquid and liquid–liquid applications. The impeller rides on a low-friction, inert bearing assembly designed to ensure low particle shedding while allowing mixing of high powder loads in large liquid volumes. All product-contacting surfaces are USP Class VI, ADCF, and 100% disposable.
Powder...
Figure 1:
Every cell line has unique nutritional requirements that must be properly balanced to achieve peak performance. However, production demands and tight timelines can make it challenging to conduct a base medium optimization for each individual cell line. To address this issue, BD has developed Select CD1000, a new, chemically defined platform medium using the proven methods of the BD AutoNutrient™ Media Design Service (AMDS) including design of experiments (DOEs) developed specifically for biopharmaceutical cell lines.
BD Select CD1000 is a high-performance base medium that has also been optimized for further supplementation to enable increased process yield during scale-up and production. In Figure 1, protein production increased when a hydrolysate was added to Select CD1000. This increase was further enhanced with the addition of a hydrolyate feed at both the shaker flask and bioreactor scale. In under three months, protein production was increased nearly 250% over the original, nonoptimized for...
Biopharmaceutical manufacturers face increased pressure to commercialize new drugs faster and at a lower cost. To meet existing and future demand and maintain a competitive advantage, many manufacturers are striving to develop increased efficiencies within their manufacturing processes. This is where single-use systems — particularly single-use transfer lines — can deliver significant value, including added flexibility, improved production yields, and increased cost savings.
The trend today is toward transfer lines or tube sets that use single-use connectors and filters to achieve sterile media transfer between process equipment or between production suites. A key benefit of single-use transfer lines is their ability to help boost productivity and accelerate time to market by reducing the downtime associated with cleaning and validation. This, in turn, helps reduce operational expenses by minimizing labor, chemical, water and energy demands. Additional cost savings stem from reduced validation efforts, gr...
To reduce scale-up costs and facility capital expenses, many biotechnology companies are transitioning their stainless steel infrastructure to single-use or disposable systems. As single-use bioreactors have matured, their market acceptance has rapidly expanded. The move to disposables is in many instances driven by reduction in sterilization and cleaning requirements, improved plant flexibility for multiproduct manufacturing, reduced product changeover costs, and faster time to market for new products. To fully enable the single-use paradigm, the automation software, hardware, and single-use sensors must be designed specifically for this application; moreover, both validation documents and leachables/extractables certifications must be readily available for cGMP implementations.
Turn-Key Solutions
Finesse provides turn-key, configure-to-order measurement and automation solutions for both single-use bioreactor types: “rocker” (e.g., GE/Wave) and “stirred-tank” (e.g., Thermo Scientific/HyClone) systems. Th...
Figure 1:
Bioprocess monitoring and automatic feeding of key nutrients such as glucose and amino acids are critical to obtain optimal high productivity in cell culture systems. Here we present a complete, closed-loop system that integrates a Groton Biosystems Automated Reactor Sample System with a nutrient analyzer and a reactor control system that demonstrates a powerful, automated online, real-time feeding system.
Materials and Methods
A Groton Biosystems Automated Reactor Sampler (ARS-M) is connected to a four-vessel DasGip Parallel Bioreactor System (PBS, Jülich, Germany) controlled by the DasGip Control 4.0 OPC Edition Software (DCS) and a YSI7100 Biochemistry Analyzer (YSI, Yellow Springs, Ohio). Data and control were transmitted between the individual units of the Integrated System by OPC protocol.
Glucose was chosen as the feedstock. The ARS-M delivered on a programmed schedule an aliquot of reactor media to the nutrient monitor. The glucose concentration data point was transmitted to the ARS-M, w...
Figure 1:
The Mobius CellReady 3-L bioreactor is a single-use, stirred-tank bioreactor ideal for bench-scale cell culture. The CellReady 3-L bioreactor is preassembled and gamma irradiated, significantly reducing turnaround times typically associated with glass bioreactors. There is no need to clean, assemble, or autoclave between experiments. The CellReady 3-L bioreactor ensures maximum operational flexibility with prefitted, weldable tubing and vent filter; two sparging options; and compatibility with most standard bioreactor controller configurations. Designed to replace traditional glass, bench-scale bioreactors, the CellReady 3-L bioreactor has additional functionality through integrated side sampling, addition, and drain ports.
The CellReady 3-L bioreactor was designed to directly replace existing 3-L glass bioreactors, which requires verification of comparable functionality as compared with glass bioreactors. The data shown here demonstrate that with regards to mammalian cell cultivation the CellRe...
Figure 1:
New Brunswick Scientific has developed three new versions of Supervisory Control and Data Acquisition (SCADA) software programs. These SCADA programs allow fermentation or cell culture processes to be automated, monitored, and controlled. The three packages — BioCommand® Track and Trend, BioCommand Batch Control, and BioCommand Batch Control Plus — provide a variety of tools and the flexibility needed for any process requirement. Whether you require only basic trending and batch reporting, programming for process automation and optimization, or security and audit trails compatible with 21 CFR Part 11 regulations, there is a BioCommand package that will meet your needs.
In this article, the advantages of using a SCADA program will be explained and the differences between the three BioCommand packages will be identified. We will review the secure event logging and audit trail features of Batch Control Plus and provide an example of a control program created using BioCommand’s intuitive control edi...
Figure 1:
Biological drugs are an increasingly important and growing part of the pharmaceutical industry but are associated with a number of challenges that have constrained their application. The lack of efficacy due to short protein half-life and/or poor availability is an issue that leads to a requirement for both high and frequent dosing (Figure 1). Such administration demands can result in unwanted side effects and limit the therapeutic benefits, as well as elevated patient dosing that creates an extra burden on manufacturing capacity and treatment costs. Albumin fusion technology (albufuse) has been designed to address these shortcomings.
Half-Life Extension Benefits
Albumin is a benign natural scaffold and carrier protein found at high concentrations in plasma with a typical half-life of 20 days. albufuse takes advantage of these properties of albumin; it is the molecular fusion of albumin to protein drug candidates providing a single hybrid protein from a contiguous cDNA for the “target” peptide o...
Quality control is very often perceived as the last step in manufacturing a product. But quality is not a consequence of quality testing; it must be built into the manufacturing process. At the beginning of the 21st century, the FDA Advisory Committee for Pharmaceutical Science (ACPS) began to address this problem, calling for consistent regulations in designing, analyzing, and controlling pharmaceutical manufacturing processes. As a result, process analytical technology (PAT) was developed providing a framework for innovative pharmaceutical manufacturing and quality assurance.
The goal of PAT is to understand and control the manufacturing process in real time (“in-process-control”). This includes the reduction of production time, the prevention of batch rejections, real-time release, and the increase of automation. Important for achieving these goals is primarily the identification of variables affecting the process, followed by the implementation of appropriate process measurements.
PAA Laboratories fol...
Figure 1:
The effect of prefiltration on the scalability of 0.1-m rated membrane filters is evaluated using a specific cell culture medium that showed reduced filtration throughput (in terms of L/m
2
) with pleated cartridges.
Biopharmaceutical process developers seek to size membrane filters more accurately to improve process economics. Understanding the scalability of filters from discs to pleated cartridges has thus become important. Contradictory claims on the scale-up between different filter configurations have been made in the literature (
1
,
2
). Recently, we reported that 0.1-µm rated Fluorodyne® EX grade EDT membrane filters showed good scalability from discs to pleated cartridges with a variety of cell culture media feeds (
3
). For some media additives, however, all 0.1-µm rated membrane filters tested, including Fluorodyne EX grade EDT membrane, exhibited nonlinear scalability (throughput scaling factor <1). In this application note, we show that for such solutions, implementation of a coar...
Figure 1:
This protein proved to be “difficult to express” in both mammalian (CHO) and yeast (
Pichia pastoris
) expression platforms. In CHO cells, expression levels were very low, <50 mg/L/day, and the product was subject to proteolysis and aberrant O-glycosylation. In yeast, expression levels were highly variable, the product was heavily mannosylated, and it was also subject to extensive proteolysis. Glycosylation is known to be immaterial to either the pharmacological or pharmacokinetic properties of this protein. Therefore, mammalian or yeast-cell expression is inappropriate because these hosts create troublesome contaminants of glycosylated products, not to mention the degraded target protein as well.
The Pfēnex team designed a strategy to address the expression of this proteolytically sensitive, disulfide-bonded protein. Multiple
sec
-type secretion leaders were fused to the coding sequence to target the growth factor to the periplasmic space. Once constructed, these expression plasmids were inser...
Figure 1:
Bioprocess fermentors are typically equipped with real-time monitoring devices such as temperature, pH, oxygen, or even viable cell density. The advantages of on-line over off-line measurements are easiness of use, timely results, and the possibility of immediate responses to the measured value.
By contrast with the well-equipped fermentors used for production, the disposables used in bioprocess development have lacked such on-line measurement devices so far. Cultivation conditions could be monitored only by means of off-line sampling, typically performed at a predefined point in time. These predefined measurement times do not take into account the metabolism of a cultivated biological system.
This situation has changed. PreSens Precision Sensing is offering noninvasive sensors for the on-line measurement of oxygen, pH, and even CO
2
in small devices such as shake flasks, spinner flasks, and microplates.
PreSens offers ready-to-use disposables with integrated sensors and the appropriate measure...
+1SAFC Biosciences® since 1971 manufactures high quality sera, cell culture media, and specialty products for commercial firms using mammalian and insect cell culture methods to develop and produce biopharmaceuticals. SAFC Biosciences focuses on organizations involved in vaccine, human therapeutics, and animal health in all stages of development from preclinical through clinical phases I, II, and III, to commercial products. SAFC Biosciences has successful relationships with pharmaceutical, established and start-up biotechnology companies, contract research and manufacturing organizations, agricultural-related firms, and nonprofit organizations.
Traditional systems involving chicken eggs and primary chick embryo fibroblasts face a number of serious limitations, including quality and quantity control, risk of contamination, and inflationary costs of serum for growing the fibroblasts. In the past decade, however, bioproduction of vaccines has undergone a renaissance as new manufacturing techniques and new ind...
Figure 1:
Deployment of new GMP biomanufacturing capacity is a battle of tradeoffs (Figure 1). Push to accelerate your timetable? Cost or quality considerations must be compromised. Need to drive down total investment? Build-out time or quality must be sacrificed. As single-use technology establishes wider acceptance, certain tradeoffs may be lessened, but serious challenges remain.
Overcoming the limitations intrinsic to conventional biomanufacturing strategies requires a fundamental rethinking of bioproduction architecture. With the FlexFactory® biomanufacturing platform and a deeply experienced team to support its effective implementation, Xcellerex offers a new level of freedom from historical limitations.
In this paper, we examine eight critical considerations facing the biopharmaceutical and vaccine production landscape, and how each is addressed with this new, proven approach to biomanufacturing.
Question #1: Can you achieve a level of process productivity suitable for commercial success?
As the bi...
Equipment automation is the key to the emerging needs of laboratory efficiency. Automation reduces the risk of manual mistakes, minimizes experimental repetitions, and saves material. This retains lower operation costs, increases efficiency, and eases factorial-designed experiment approaches, such as design of experiments (DoE). Automated laboratory processes combined with comprehensive data management programs open the way into laboratory- and site-spanning information networks.
With its Parallel Bioreactor System, DASGIP supports laboratory automation in microbiology and cell culture: It offers excellent opportunities for automation of cultivation processes at small scale including the integration of third-party equipment. Open communication with supervisory control systems, LIMs, and data historians ensures interoperability with existing information networks.
Process Automation Supports PAT
Keeping in mind regulatory demands such as the Process Analytical Technology initiative (PAT) at R&D level can he...
As the pharmaceutical industry focuses on developing biological drugs derived using cell culture processes, the ability to optimize the growth, health and potential yield of the cell line becomes increasingly important. Finding the optimal cell culture media is a daunting and time-consuming process due to the amount of potential media components and the unique requirements of the cell line being researched. Osmolality information is a crucial component to this research because of its close relationship between cells and media.
Background
Most current osmometers use freezing point depression technology to measure the total solute concentration of a liquid solution. Freezing point is considered the gold standard method in the pharmaceutical industry due to its accuracy and its broad applicability for almost any liquid sample type including complex media mixtures. Historically, high-throughput osmolality determinations have been limited due to the ability of the systems to process only one sample at a time r...
Bioengineering AG looks back at over 35 years of experience in the planning and construction of biotechnological plants. Holistic solutions have become the status quo: from inoculation to harvest, from feed line to CIP-system, and from automation to validation. Because we do not want to limit our know-how to customers fermenting with Bioengineering equipment, we made components for all process steps available as single parts. To support progress in biotechnological processes, we also develop custom-made components for special applications.
Operational Safety
Crevices, deadlegs, and leaks are potential sources of microbial contamination or environmental hazards in biotechnology. Valves and sterile couplings are necessary for process control and maintenance, and often constitute the transition from unsterile to sterile plant segments. Bioengineering manufactures sterile couplings with deadleg-free passage and seals flush with the pipe wall. Hygienic diaphragm valves are available from manually operated one-...
Figure 1:
The bacterial endotoxins test (BET) is required for most parenteral drugs and medical devices that come into contact with the human bloodstream.
Limulus
amebocyte lysate (LAL) derived from horseshoe crabs is used for the BET, and several methodologies have been developed. Popular quantitative LAL techniques are the kinetic turbidimetric assay (KTA) and the kinetic chromogenic assay (KCA). It is, however, well known that the endotoxin standards used in both those assays are profoundly influenced by the techniques and materials used by trained LAL analysts each and every day. The type of pipettes used, the type of dilution tubes used, the length of time used to vortex endotoxin standards, and the stability of the liquid stock endotoxin solution are variables that laboratory personnel manage every day. The Portable Test System (PTS) is a new FDA-approved KCA method that uses an archived standard curve. Here, advantages of the archived standard curve method are demonstrated and discussed.
Material...
ELISA-based sandwich assays have been a traditional workhorse across the drug discovery continuum for many years, but they have significant limitations. Gyros® offers a more efficient solution for performing high-throughput, automated immunoassays on a miniaturized, fluorescence-based platform.
A head-to-head comparison of the microfluidics-based Gyrolab® technology and ELISA demonstrates clear distinctions:
What Is Gyrolab?
Gyrolab is a bioanalytical, automated, fluorescence-based system that employs microfluidics technology to perform immunoassays at nanoliter scale. At the core of the Gyrolab system is a compact disc (CD) microlaboratory, in which are embedded microstructures containing affinity capture columns. By integrating microfluidic CD technology with sensitive detection methods and intuitive software, this automated system yields quantitative assay data and highly reproducible results.
How Does It Work?
Designed based on microfluidic flow-through principles, Gyrolab’s CD-based microlaboratory a...
In industries where implementation of process analytical technology (PAT) is bringing significant benefits and improvements to many processes, any new supporting technology that helps manufacturing efficiency and lowers production cycle time is often adopted in various functional areas throughout the organization. Bioprocessing operations often need to determine nucleic acid or protein concentrations reliably and with fast sample turn-around time at various stages during manufacturing.
A common practice today is UV-Vis absorbance spectroscopy using conventional cuvette-based spectrophotometers. However, a novel approach using a microvolume UV-Vis analysis method is providing significant improvements in processing time as well as reliability.
The Thermo Scientific NanoDrop™ 8000 UV-Vis spectrophotometer uses a patented sample retention technology that allows direct measurements for samples as small as 1 µL across a broad concentration range without the need for dilutions. Analytical laboratories, manufactu...
