cell culture – BioProcessing Journal

Outsourcing Clinical Trial Material Manufacturing: Regulatory and Quality Issues for the Twenty-First Century Biotechnology Product

The United States Food and Drug Administration (FDA) considers antibodies and recombinant proteins as “well-characterized products.” This is based on FDA’s comfort level with reviewing multiple products over an extended period of time. This designation relates to the product, not necessarily to the system that is used to manufacture the product nor to the facility where the product is manufactured. The initial guidance document was published in 1995, prior to the use of other systems and when the majority of products were still based on mouse hybridoma technology that was 20 years old…

Biologics Production

antibodies biopharma guidance documents biopharmaceutical cell culture clinical trial material FDA mouse hybridoma quality issues recombinant protein well-characterized products

Expanding Horizons: Opportunities in Eastern Europe for Contract Research Organizations

The contract research organization (CRO) industry is rapidly adapting to the exigencies of pharmaceutical and biotechnology companies. For a start, the function of CROs is no longer confined to offering only clinical trial outsourcing services. Instead, many CROs now provide a full complement of services that guide products from inception to commercial implementation…

Biologics Production

biopharmaceutical biopharmaceutical research organization cell culture contract research organization cro pharmaceutical

Hybridoma and CHO Cell Culture Using the New Brunswick™ S41i, An Environmentally-Friendly, “Low Emission” Incubator Shaker

Automobiles contribute significantly to CO2 emissions in the atmosphere. However, laboratory equipment such as CO2 incubators can potentially release over 120,000 liters of CO2 gas per year. The “epGreen initiative” established by Eppendorf® will ensure that their products are developed with the mindset of reducing environmental impact…

Biologics Production

carbon dioxide cell culture cho cells co2 environmental compact hybridoma culture

Adventitious Virus Contamination Testing: Massively Parallel Sequencing in a Multimodal Solution for Biopharmaceutical Safety Testing

Virus contamination of commercially valued cell cultures can be a health risk to the general population and imposes financial burdens on manufacturing and biopharmaceutical companies. We investigated the use of massively parallel sequencing/next-generation sequencing (MPS/NGS) and bioinformatics for the detection and subsequent identification of adventitious contamination. Specifically, the Illumina® HiSeq 2000 instrument, in the 2 × 100 base pair (bp) paired end (PE) run configuration, was used to determine the identity and limit of detection of a DNA virus spiked into a virus vaccine, and an RNA virus spiked into a mammalian master cell bank (MCB). This configuration provided sufficient sequence read lengths and depth of coverage to detect and identify spiked-in SV40 and measles viruses in a background of vaccine, MCB, and host nucleic acid that make up the bulk of these samples. Furthermore, the detection of 30 SV40 reads within one tested sample suggested a < 1 plaque forming unit (pfu) sensitivity in a background of 2.8 × 107 infectious adenovirus serotype 5 (Ad5) particles. Results from subsequent virus vaccine testing suggested the presence of non-viable virus DNA contamination in the sample. Therefore, we propose that a multimodal approach, in which broad-range screening for known or unknown adventitious agents by MPS/NGS is complemented by targeted virus detection assays (e.g., PCR-based and infectivity assays), should provide the most useful safety monitoring information. We suggest that MPS/NGS and the accompanying bioinformatics is a sensitive, broad-range, and long-lasting tool with the ability to improve upon existing biosafety testing within a larger testing program…

Biologics Production

bioinformatics cell culture dna contamination host cell proteins mammalian cell bank massively parallel sequencing master cell bank mcb measles virus next generation sequencing PCR plaque forming unit SV40 vaccine production vaccines viral contaminants virus contamination virus detection assays virus spikes

Virus Clearance in Your Process from Start to Finish

The safety of biological products (biologics) derived from in vitro cultured cell lines of animal origin is dependent both on clinical studies to evaluate efficacy, and a matrix of controls throughout the manufacturing process to assure consistency, quality, and safety of the marketed product for human use. One major area of concern is virus safety. Through the combination of: (A) careful selection of raw materials; (B) testing of process intermediates; and (C) virus clearance evaluations of the individual steps in the manufacturing process; biologics manufacturers can demonstrate that their products are free from detectable adventitious and endogenous viruses. Comprehensive regulatory guidance suggests approaches for virus testing of biologics at early and later stages of clinical development and, although some countries have specific requirements, many of these approaches are harmonized worldwide. In general, regulatory authorities expect purification processes to include multiple steps with complementary, or orthogonal, methods for virus reduction including inactivation and separation or removal. Each step in the process may be more or less effective for reducing levels of different test viruses, and therefore selection of the appropriate panel of test viruses for the specific product is critical for viral clearance studies…

Biologics Production

biologics biopharmaceuticals cell culture mammalian cell culture raw materials purity viral clearance viral safety testing virus safety

Rapid and Effective Monitoring of Baculovirus Concentrations in Bioprocess Fluid Using the ViroCyt® Virus Counter®

The licensing of recombinant vaccines produced using the baculovirus expression vector system (BEVS) has cleared the way for the production of a variety of biopharmaceuticals produced using this technology. Obtaining accurate estimates of both total and infectious baculovirus titer in upstream and downstream bioprocess fluids is one of many process controls that will need to be addressed during the development phase of a product’s lifecycle. Traditional plaque-titer methods require 5–7 days of incubation in order to reveal plaques that may be enumerated, and is further complicated by plaques created by multiple viruses that may be scored as a single plaque, thereby lowering the titer estimate. Titer assays based on polymerase chain reaction (PCR) have been developed, but they measure the presence of baculovirus genes, not virus particles. This often results in titers one or two logs higher than the actual titer. Immunoassays correlate with host cell infection and virus replication, but they too can be time-consuming and difficult to interpret. Our goal was to identify a method that would provide estimates of both total and infectious virus particles in as close to real-time as possible. We have evaluated the ViroCyt Virus Counter over the course of three years and have found it to provide accurate and reproducible estimates of both titer types in as little as 30 minutes. We have created an algorithm that converts total virus particle counts into estimates of infectious titer and tested these values in virus amplifications. The Virus Counter method of titer determination has also been used to track the quantity of virus particles in the culture supernatant of stirred-tank bioreactors infected with standard baculovirus stocks and with baculovirus-infected insect cells (BIIC)…

Biologics Production

baculovirus express system baculovirus expression system baculovirus titer baculovirus vector BEVS biologics biopharmaceutical manufacturing biopharmaceuticals cell counting cell culture cgmp immunoassays insect cell culture recombinant vaccines vaccine production viral titer virocyt

Optimising Vaccine Process Scale-Up of Attachment-Dependent Cells Using Micro Bioreactors and Microcarriers

The trend for increased vaccine production is being driven by the requirement to produce affordable prophylactic vaccines for use in emerging markets, and also for newer types of therapeutic vaccines to treat an ever-increasing array of diseases such as cancer. These drivers, coupled with the current need to produce vaccines rapidly for pandemic threats and seasonal influenza prevention, has lead to the investigation of rapid method development for optimising the scale-up of cGMP-compliant manufacturing processes…

Biologics Production

attachment-dependent cells biologics biopharmaceutical manufacturing cell culture cgmp microbioreactor microcarriers pandemic threats process scale-up therapeutic vaccines vaccine production

Which Factors to Consider When Selecting an Analytical Method for Cell Culture Fermentation: A Comparison of Four Different Metabolic Analyzer Instruments

Mammalian cell culture processes require an in depth understanding of inputs and outputs in order to maximize productivity, efficiency, and product quality. Daily monitoring of essential metabolites, nutrients, and protein titer using at-line analyzers are the building blocks of drug manufacturing process development and characterization. In order to better understand how these instruments perform, a comparative analysis was conducted using two different classes of metabolicsensing technologies, membrane-based technology (MBT) and absorption photometric-based technology (APBT). Four commercially available instruments were examined using various samples and maintenance conditions in order to emphasize the following testing criteria: specificity, linearity, range, accuracy, and precision. Samples included standard solutions with known metabolite concentrations, cell culture supernatant, and supernatant liquid spiked with additional metabolite solutions. The results from our testing indicated that the photometric analyzers yielded more accurate and consistent results than the membrane-based analyzers and were easier and less time-consuming to maintain. Ultimately, these studies summarized the capabilities and limitations for both types of analyzers and provide a critical summary for instruments used in everyday bioprocess monitoring…

Analytics Biologics Production

adsorption photometry apbt cell culture cell culture fermentation mammalian cell culture matabolic analysers mbt membrane-based technology metabolicsensing technologies

Coming ‘Round to Spheroid Culture

Cells cultured in 2D can differ in terms of both physiology and cellular responses compared with cells in vivo. This has led to a surge in the popularity of using 3D culture techniques as mounting evidence suggests that culturing cells in 3D is more representative of the in vivo environment, even to the extent that the gene expression profiles of cells from 3D cultures more accurately reflect clinical expression profiles than those observed in 2D cultures. 3D culture offers the potential for more accurate models of drug delivery and efficacy, as well as numerous clinical and research applications, and is becoming increasingly capable of integrating into high-throughput activities. Spheroids, or sphere cultures, have become an especially exciting area of 3D in vitro culture due to their great potential for use in studies that investigate growth and function of both malignant and normal tissues. These sphere cultures have contributed considerably to our knowledge of cellular responses thanks to the accuracy with which they reflect the in vivo system.

Biologics Biologics Production Cell Lines Pre-Clinical Development Research

2d cells 3d cells cell culture cho cells spheroids

Fetal Bovine Serum: Risk Management

Safety is typically viewed, perhaps unconsciously, as the result of a collection of factors, conditions, or behaviors. For example, consider “safety” in the context of personal, financial, or travel. With each, safety is defined as a set of component risks that have been managed to satisfactory levels for a particular situation. The same is true for product safety and risk, whether it be for raw materials or finished goods. The “safe” use of fetal bovine serum (FBS) is achieved by the management of controllable risks to a level that is acceptable for each particular application. For example, risk reduction requirements for research applications are not as stringent as for diagnostic, therapeutic, or manufacturing applications. Each end-user must decide on the level of risk reduction that is appropriate for their application…

Biologics Biologics Production Bioreactor Scale-Up Cell & Gene Therapy Cell Lines Fed-Batch Bioreactor Process HEK293 Mammalian Cell Culture Manufacturing Regulatory Viral Reference Materials Viral Vectors

animal sera animal-derived materials biologics raw materials biotherapeutics cell culture cgmp fbs fetal bovine serum gamma irradiation geographical origins isia safe use serum serum traceability vaccines

Tag: <span>cell culture</span>

SEARCH