BioProcessing Journal

Eco-Friendly Red Pigment Production by Sporosarcina aquimarina

Acetyl-4, 4′-diapolycopene-4, 4′-dioate, a C30 carotenoid and secondary metabolite, was produced by the Sporosarcina aquimarina bacteria using a 5.0 L fermentation vessel with a 3.0 L working volume. In the presence of tryptone, the biosynthesis of acetyl-4, 4′-diapolycopene-4, 4′-dioate production using a batch fermentation process was further improved. Production parameters like carbon source, pH, and temperature were studied, and maximum product was achieved, up to 1.2 g/L, where the secondary metabolite yield was 0.07 g/L and productivity, 0.00833 g/L/h. The organic constitution and significant red color intensity of the acetyl-4, 4′-diapolycopene-4, 4′-dioate molecule can be used in the textile industry as a dye, and a coloring additive in processed foods and pharmaceuticals.

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

bacillus thuringiensis bioinsecticide mab mir162 monoclonal antibody mouse mabs pab polyclonal antibody transgenic corn vip3aa20 protein

Monoclonal Antibody Generation and Characterization for Vip3Aa20 Protein Quantification in Transgenic Corn Plants

Numerous standardized techniques for detection and quantification of proteins are based on polyclonal antibody (pAb) use. However, because pAbs are a heterogeneous mixture of antibodies, there is the possibility of non-specific interactions or cross-reactions with non-related proteins, which is a disadvantage in the detection and quantification of target proteins. Therefore, the main objective of this study was to generate and characterize monoclonal antibodies (mAbs) for quantifying the Vip3Aa20 protein of Bacillus thuringiensis (Bt) expressed in event MIR162 transgenic corn plant.

bacillus thuringiensis bioinsecticide mab mir162 monoclonal antibody mouse mabs pab polyclonal antibody transgenic corn vip3aa20 protein

Improving Process Economics When Developing a Vaccine Portfolio

Over the last few years, the challenges of vaccine development have created perhaps an unprecedented level of scrutiny, not just within the biotech industry, but also in the consciousness of the general public. This was certainly the case during the recent H1N1 influenza outbreak. The demand to know when a vaccine would be available, and if producers could meet the global demands consistently made front page news. The challenge of rapid and scalable manufacture is of course nothing new in biopharmaceutical development and in many respects, monoclonal antibodies are leading the way as the industry moves towards the required level of industrialization.

antibody production cho cells clinical development cmo complex biologicals contract manufacturing FDA influenza novel vaccines phase 1 phase ii process economics regulatory guidelines risk-based-approach scalable manufacturing vaccine portfolio vaccines

Quality Risk Management Application Review in Pharmaceutical and Biopharmaceutical Industries

An interesting situation occurred ten years ago in an industry where quality had been the critical factor in management decision-making. Quality was abruptly bumped to second place behind pricing, which started a global rush to find lower-cost suppliers and eventually resulted in massive outsourcing. During this evolution, the importance of quality systems for the pharmaceutical and biopharmaceutical community was recognized by regulatory agencies that began establishing risk management standards.

biologics biopharmaceuticals cgmp fmea GMP iso pharmaceuticals QRM quality risk management risk analysis risk evaluation risk management

Novel Automated Micro-Scale Bioreactor Technology: A Qualitative and Quantitative Mimic for Early Process Development

With increasing time pressures to move biological therapeutics into the clinic, bioprocessing development studies have to be limited. Currently, core studies typically involve the use of shake flasks and benchtop bioreactors to select the most productive clones, optimum media, and bioprocessing conditions. The capacity for using benchtop bioreactors is especially limited as it is resource-intensive and has high capital equipment and infrastructure costs. Consequently, scientists frequently cannot perform full design-of-experiments (DoE) and are generally only able to take one or two of their most promising clones forward for partial DoE runs in benchtop bioreactors.

ambr batch culture benchtop bioreactor cell culture design-of-experiments micro-scale bioreactor miniaturised bioreactors miniaturized bioreactors

Development of a Scalable Fed-Batch Bioreactor Process for High-Titer Production of Lentiviral Vector Using an Inducible HEK293 Producer Cell Line

The rapidly growing interest for cell and gene therapies demands the development of robust, scalable, and cost-effective bioprocesses for viral vector production. For the production of lentiviral vector (LVV) at high titers, we have developed an inducible packaging system in suspension HEK293 cells from which we can also generate stable producer cell lines, in serum-free conditions. To evaluate the potential of this platform, we have generated a stable cell line that produces an LVV encoding a green fluorescent protein (GFP) and obtains 10E+07 to 10E+08 transduction units (TU)/mL at the 4 L, 10 L and 50 L scales. Functional LVV titers were maintained across all scales in bioreactors with different configurations and geometries indicating process robustness. Further, the addition of 10% feed increased the volumetric productivity by 3.5-fold in comparison to batch production, making our platform suitable for large-scale LVV production and showing a real potential for commercial manufacturing.

batch bioreactor scale-up fed-batch HEK293SF lentiviral vector LVV-RM stable producer clone

Quality Risk Management (QRM): Part I – Identifying, Evaluating, and Mitigating Threat Risks to Biopharmaceutical Enterprises

The FDA’s ICH Q9 quality risk management (QRM) guidance material is the foundation for understanding and evaluating patient risks associated with developing and manufacturing pharmaceuticals. This three-part paper describes approaches a team of subject matter experts (SMEs) can use for implementing two important applications of QRM. Part I provides a method for identifying and remediating threat risks that may affect the product’s quality or other important aspects of a manufacturing enterprise’s lifecycle, from product research and development to commercial manufacturing. The second QRM application covered in Part II manages patient risks by identifying, evaluating, and managing risks associated with process parameters (PP) on the product’s critical quality attributes (CQAs). The final paper, Part III, describes an approach for accepting or further mitigating the risks evaluated by the QRM exercise…

Biologics Production Guidance Mammalian Cell Culture Quality Risk Management (QRM) Regulatory Unit Operations

biopharmaceutical commercial biomanufacturing CQA critical quality attribute enterprise lifecycle mark f. witcher PP process parameters QRM quality risk management remediating threat risks

Low-Pressure Liquid Chromatography Using the EcoPrime® With Enhanced Buffer In-Line Dilution

Chromatography and buffer in-line dilution (BID) are integrated on the same low-pressure liquid chromatography (LPLC) system combining two unit operations into a single, space-saving platform. The EcoPrime’s BID option will significantly reduce buffer storage and lower operating expenses associated with buffer preparation…

Biologics Production Buffer Formulations Chromatography Low-Pressure Liquid Chromatography (LPLC) Manufacturing Process Automation Unit Operations

automated buffer prep buffer storage EcoPrime lower operating expenses LPLC Process Automation space saving unit operations

NGS-Based Viral Safety Testing for ATMPs: Mandatory or Just an Option?

The heterogenous group of advanced therapy medicinal products (ATMPs) are biologics with frequently limited viral safety profiles. As compared to well-established biologics such as monoclonal antibody products, the risk of virus contamination is significantly higher for some ATMPs. The standard approaches and tools used to mitigate the viral risk have limitations, leaving open the chances of missing virus contamination in an ATMP manufacturing process in both upstream and downstream. Next-generation sequencing (NGS) technology can overcome the residual risk by having the potential to detect any kind of virus contamination based on its inherent capability to detect any kind of nucleic acid in a sample. It perfectly combines the benefits and compensates for the downsides of the existing testing tools. It will replace a bunch of different established testing methods at improved turnaround times and, in the end, reduced overall costs. The combination of these characteristics is making NGS-based virus testing an in-demand and preferred approach to mitigating the virus contamination risk across all kinds of biologics mid- and long-term.

Bioinformatics Biologics Production Cell & Gene Therapy Regulatory Risk Analysis and Management Viral Reference Materials Viral Vectors

advanced therapy medical products advanced therapy medicinal product ATMP next generation sequencing next-generation sequencing NGS NGS-based virus testing nucleic acid sample release testing viral contamination viral risk mitigation viral safety profiles virus safety testing

Practical Considerations in Using an Equivalence Approach to Establish Lot Release Limits for Vector Dose

To demonstrate that a dose-determining assay is fit for purpose, the measurement uncertainty associated with a reported release test result must be suitably small. The establishment of a corresponding product specification is inextricably linked to the tolerance for error in assigning a dose value for a vector lot. By adopting an equivalence-based lot release model which includes a total error approach to assay qualification, specific testing strategies can be evaluated quantitatively for dose error and lot release decision risks throughout the drug development process. This article aims to reinforce how the concepts tied to an equivalence-based lot release model are interrelated and applied in practice. It provides in-depth explanations of fundamental concepts and clarifies common misunderstandings for quality control, quality assurance, and regulatory affairs personnel held accountable for decisions made in vector dose assignment and product lot release.

Risk Analysis and Management Viral Vectors

coefficient of determination confidence interval consistency dosage dose-determining assay lot release measurement uncertainty quality assurance

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