BioProcessing Journal

PCR Turns 25: Celebrating the Past and Anticipating the Future

Beginning with the production of the very first recombinant deoxyribonucleic acid (DNA) molecule in 1972 and continuing through the following decade, DNA-focused molecular biology researchers firmly positioned themselves on the cutting edge of scientific inquiry. With each passing day, month and year, these researchers made significant leaps forward in understanding, modifying and manipulating the key roles that DNA plays in all living organisms. Some of the key milestones…

Nucleic Acids Research

amplification gene expression genetic research genotyping molecular research PCR polymerase chain reaction

Reflections On the GFP Nobel “Buzz”

In October 8, 2008, the Nobel Prize for Chemistry was awarded to Osamu Shimomura, Martin Chalfie, and Roger Tsien ostensibly for the discovery, cloning, and applications of green-fluorescent protein (GFP). This protein is associated with bioluminescence in coelenterates, including jellyfish and sea pansies. This award is as much a celebration of basic research as it is an acknowledgement of the achievements of three accomplished researchers…

Research

bioluminescence capstone terminal cloning chromatofocusing green-fluorescent protein research funding technology transfer

Rapid Development of High-Performance Stable Mammalian Cell Lines for Improved Clinical Development

Since the first approval for human use of a recombinant protein therapeutic, this sector of the pharmaceutical market has grown rapidly. The first approved protein therapeutics were small, non-glycosylated proteins such as insulin and human growth hormone; they were produced in bacterial systems. With the advent of mammalian cell-based production systems, it became possible to produce more complex, glycosylated proteins for use as recombinant therapeutics…

Biologics Production

cell line selection mammalian cell line development recombinant proteins stability therapeutic proteins vector construction

Manufacture and Stability of the Recombinant Adeno-Associated Virus Serotype 2 Vector Reference Standard

Recombinant adeno-associated viral (rAAV) vectors have proven to be efficient vehicles for gene transfer in animal models. The attractive features of this vector system are long-term gene expression with little or no associated toxicities following administration to a variety of tissues. Previous and ongoing clinical trials in humans demonstrate a very good over-all safety profile. However, one of the caveats of this work that has been carried out by several laboratories is the inability to normalize vector doses administered by different investigators to animals and humans. Most of the work to date has involved AAV serotype2 vectors, but vector systems based on other AAV serotypes are being rapidly developed…

Viral Reference Materials Viral Vectors

adeno-associated viral vectors FDA helper virus-free transient transfection infectious titer purification stability testing standards

Expression Vector Engineering for Cell Line Development — New Roles for “Old” Sequences

Expression vector and cell line engineering is the basis for expression and industrial production of biopharmaceuticals. The ultimate goal is to obtain clonal cell lines that secrete the protein of interest with high cell-specific productivity, and at consistently high levels over an extended number of cell generations, allowing for scale-up and cost-efficient large-scale manufacturing. Productivity and stability of expression are thus the prerequisites for developing commercially viable processes…

Manufacturing

cell line development expression vector engineering intervening sequences protein expression transgene expression

Optimization of Peptone Supplementation In the Development of a High-Performance CHO Growth and Production Medium

Optimization of the key factors in a biopharmaceutical process is necessary to meet the continuous rise in the production demands. One of the key factors in the process addresses the diverse nutritional and growth requirements of the cell culture—peptone supplementation. Peptones are low molecular weight protein digests, which provide nutrients such as amino acids, peptides, vitamins, and minerals to the culture medium. They are widely accepted as the supplements that enhance the performance of a chemically-defined cell culture medium, and have successfully been used for over thirty years…

Biologics Production

animal-free peptones cell culture media media supplement peptone supplementation process optimization

Membrane Chromatography: An Alternative to Polishing Column Chromatography

With the development of bacterial fermentation and mammalian cell culture as the sources for new recombinant products came a standardization of raw feed stocks. Therefore, manufacturers came to share the same types of problems. This standardization allowed a more systematic approach to process development divided into upstream (bacterial and yeast fermentation or mammalian cell culture) and downstream processing activities…

Manufacturing

antibody purification chromatography downstream processing membrane chromatography purification viral clearance

Extreme Pipetting Mission #2: Holy Geyser! Understanding How Pipetting Warm and Cold Liquids Affect Data Integrity (part 2)

After braving the home of the world’s worst weather to study the affect of barometric pressure on pipette volumes, the ARTEL Extreme Pipetting Expedition team embarked on its next mission to explore the impact of another environmental condition on laboratory data. The destination: Yellowstone National Park, boasting the world’s largest active geyser field, and more than 10,000 geysers, steam vents and mud pots. The ARTEL team landed in the 2.2 million acre park and realized they were certainly not in Mount Washington, New Hampshire anymore (the venue for Mission #1). As hot springs bubbled and a geyser spewed steam up to 135 feet in the air just a short distance from Expedition members, the thermal variation at Yellowstone National Park was evident…

Research

data integrity pipetting thermal disequilibrium volume variation

How to Construct a Monoclonal Antibody Factory: A Comparison of Production Costs in Fed Batch and Perfusion Culture with Microcarriers (part 2)

A basic engineering study has been performed to evaluate three different strategies for the production of monoclonal antibodies (MAbs) from Chinese hamster ovary (CHO) cells. Cells are expanded in suspension culture and are then inoculated into either fed batch or perfusion culture for MAb production. The first strategy, which is also the current industry standard, uses fed batch culture with the cells in suspension in a stirred tank fermenter. The second strategy uses perfusion culture with the cells immobilized on Cytopore™ microcarriers in a stirred tank fermenter. The third and final strategy is perfusion culture with Cytoline™ microcarriers in a fluidized bed fermenter. Perfusion cultures, while leading to a somewhat lower product titer, were characterized by a much smaller equipment footprint. This in turn led to a >30% reduction in investment costs and a 12% reduction in MAb production costs calculated over five years of depreciation and ten years of production time…

Manufacturing

continuous perfusion cost reduction facility design fed-batch microcarrier perfusion technology

Current Practices in Heart Valve Preservation (part 2)

Storage and shipping variables may significantly impact cryopreserved product quality. In this particular article, the focus is on temperature control issues. The AATB (American Association of Tissue Banks) Standards for Tissue Banking state that cryopreserved cardiac allografts shall be maintained at temperatures of –100°C or colder. Heart valves are usually stored below –135°C in vapor phase nitrogen. There have been few published studies of higher (warmer) storage temperatures. Most heart valves are transported using dry shippers that maintain vapor phase nitrogen temperatures. These containment devices are expensive, and the costs for two-way shipping are significant due to their size and weight. Considerable savings could be had if dry ice shippers with temperature excursions warmer than the AATB standard (of up to –70°C) could be employed…

Cell & Tissue Banking

cardiac medicine cryopreservation human valve allografts preservation storage temperature-dependent changes

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