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An Overview of Metal–Peptide Complexes Used in Biomedical Research

by Archana Gangakhedkar
Volume 12, Issue 2 (Summer 2013)

Metal peptide complexes play an important role in understanding the mechanisms of complex diseases at a molecular level. Peptides are good chelators for metal ions and can form important tools in the drug and medical device industries. Transition metal ions are characterized by their ability to assume different oxidation states and serve as centers for metalloenzymes that catalyze electron transfers. Some transition metals act as Lewis acids and become an active site on enzymes and also catalyze reactions (e.g., peptidases, phosphatases). Metal peptide complexes are involved in Alzheimer’s disease (AD) and are used as cancer diagnostics. Many biomolecules (molecules produced by living cells) like metabolites, hormones, and vitamins are capable of forming strong, multidentate metal ion complexes. Species in the plant and animal kingdoms synthesize specific metal ion chelators that bind to essential metals with extremely high affinities. Plants and animals including humans possess a number of unique biomolecules that can coordinate metal ions. Peptide conjugation is a widely used and effective method for improving both cellular and nuclear entry of a variety of molecules...

Citation:
Gangakhedkar A. An Overview of Metal–Peptide Complexes Used in Biomedical Research. BioProcess J, 2013; 12(2): 47-51.
http://dx.doi.org/10.12665/J122.Gangakhedkar.

Posted online August 1, 2013.

 
Rapid Manufacture and Release of a GMP Batch of Avian Influenza A(H7N9) Virus-Like Particle Vaccine Made Using Recombinant Baculovirus-Sf9 Insect Cell Culture Technology

by Timothy J. Hahn, PhD, Denise Courbron, Mervyn Hamer, Malek Masoud, James Wong, Konnie Taylor, Jody Hatch, Mike Sowers, Erica Shane, PhD, Margret Nathan, Hua Jiang, PhD, Ziping Wei, PhD, John Higgins, Kwan-Ho Roh, John Burd, Diana Chinchilla-Olszar, PhD, Monique Malou-Williams, Daniel P. Baskind, and Gale E. Smith, PhD
Volume 12, Issue 2 (Summer 2013)

Novavax reported that, in 28 days following public release of the avian influenza A/Anhui/1/2013 (H7N9) virus gene sequences, its recombinant DNA and baculovirus-Spodoptera frugiperda cell culture-based technology was used to produce a virus-like particle vaccine to avian influenza A(H7N9) virus, and murine animal challenge studies were initiated. This report describes Novavax’s manufacturing process and the coordinated timing of critical activities necessary to produce and release a clinical batch of avian influenza A(H7N9) virus virus-like particle vaccine, under current good manufacturing practices, within three months from the time that the virus genomic sequences for this potential pandemic influenza virus were reported...

Citation:
Hahn TJ et al. Rapid Manufacture and Release of a GMP Batch of Avian Influenza A(H7N9) Virus-Like Particle Vaccine Made Using Recombinant Baculovirus-Sf9 Insect Cell Culture Technology. BioProcess J, 2013; 12(2): 4-17.
http://dx.doi.org/10.12665/J122.Hahn.

Posted online July 29, 2013.

 
Five Insurance Tips for Biotechnology Companies Facing Product Recalls or Liability Claims

by Jonathan M. Cohen
Volume 12, Issue 2 (Summer 2013)

Biotechnology companies — those that research and manufacture products through the use of biological techniques such as genetic engineering and the development of specialized strains of biological substances — constitute an increasing segment of the US economy. These companies might: (a) create a new type of insect- or drought-resistant corn by the modification of genes; (b) alter naturally occurring enzymes to aid in manufacturing or to help produce foods; (c) use recombinant DNA to create medicines that are remarkably effective in curing or treating disease; or (d) use any of a number of other techniques to create beneficial and potentially lucrative products. Just like other industries, biotechnology groups can be faced with recalls of their products, either voluntary or government-mandated, or claims that their products have caused bodily injury or property damage to their customers. Because biopharmaceuticals are produced by — or extracted from — a biological source, the chances of a product recall are higher than that of a synthesized drug. In April 2012, a report by GBI Research, an independent research firm, concluded that biologics were involved in more recalls, voluntary or mandatory, than drugs from other sources for the four-year period from 2007 through 2010...

Citation:
Cohen JM. Five Insurance Tips for Biotechnology Companies Facing Product Recalls or Liability Claims. BioProcess J, 2013; 12(2): 43-46.
http://dx.doi.org/10.12665/J122.Cohen.

Posted online July 24, 2013.

 
Effective Site-Specific Isotopic Labeling: Expression Optimization Using Specialized Media

by Kenneth C. Bonanno
Volume 12, Issue 1 (Spring 2013)

For decades, recombinant genes have been expressed in a variety of cellular systems to generate protein reagents that are the potential targets for new small-molecule drugs. As these targets become more complex, researchers have developed innovative methods to study the structure of these proteins and their interactions with potential drugs. BioExpress® 2000 media (Cambridge Isotope Laboratories, CIL) incorporates isotope-labeled amino acids into recombinant proteins expressed in baculovirus-infected insect cells to assist nuclear magnetic resonance (NMR) structural studies. In order to use this labeled media most efficiently, expression conditions must be optimized at a small-scale prior to production...

Citation:
Bonanno KC. Effective Site-Specific Isotopic Labeling: Expression Optimization Using Specialized Media. BioProcess J, 2013; 12(1): 43-46. http://dx.doi.org/10.12665/J121.Bonanno.

Posted online May 30, 2013.

 
Bovine Serum Albumin Partitioning in Aqueous Two-Phase Systems: Effects of Variables and Optimization

by Nangarthody Sindhu, Sivakumar Kalaivani, and Iyyaswami Regupathi
Volume 12, Issue 1 (Spring 2013)

The objective of this study was to optimize process conditions for the effective partitioning of bovine serum albumin (BSA) using response surface methodology (RSM). Initially, four different salts (tripotassium citrate, tripotassium phosphate, sodium carbonate, and sodium sulphate) were tested for the ability to partition BSA. Among the salts chosen, tripotassium citrate was observed to yield a high partition coefficient. The effect of phase forming components: concentration, PEG molecular weight, and pH were studied for a PEG/tripotassium citrate system and the information obtained was utilized to fix the ranges in RSM studies. Four different independent variables (PEG 2000, tripotassium citrate, NaCl concentrations, and pH) were considered for RSM studies and the responses generated were partition coefficient (k) and percentage yield. A statistical model was developed and the values obtained were 99 % within the confidence level. Optimal conditions of the system were found as: 0.25 M sodium chloride, 32 % PEG 2000 (w/w), 16 % tripotassium citrate (w/w), pH 6.0, a partition coefficient of 6.03, a recovery of 91.76 %, and a controlled operating temperature of 303.15 K...

Citation:
Sindhu N, Kalaivani S, Regupathi I. Bovine Serum Albumin Partitioning in Aqueous Two-Phase Systems: Effects of Variables and Optimization. BioProcess J, 2013; 12(1): 29-41. dx.doi.org/10.12665/J121.Regupathi.

Posted online May 30, 2013.

 
A Comparative Bioreactor Vessel Study: Conventional Reusable Glass and Single-Use Disposables for the Production of Alkaline Phosphatase

by Taylor Hatton, Shaun Barnett, Ma Sha, PhD, and Kamal Rashid, PhD
Volume 12, Issue 1 (Spring 2013)

Single-use, stirred-tank bioreactor systems have been used in large-scale production for a number of years. Bench-scale, stirred-tank bioreactors have not been commercially-available for single-use until recently. The New Brunswick™ CelliGen® BLU pitched-blade bioreactor was introduced in 2009, and the CelliGen BLU packed-bed bioreactor, in 2012. Little information is currently available on the utility of these bioreactors for bench-scale production of recombinant products. Thus, we designed this study to perform multiple comparisons with these single-use bioreactors and their traditional glass vessel counterparts. The data comparisons included: (1) CelliGen BLU pitched-blade vs. glass pitched-blade; and (2) CelliGen BLU pitched blade in batch mode vs. CelliGen BLU packed-bed in perfusion mode. Chinese hamster ovary (CHO) cells were used to measure alkaline phosphatase (ALKP) production in each bioreactor. The final measured concentration of ALKP, after eight days of batch-mode culture in the single-use, pitched-blade bioreactor, was 1.6 U/mL compared to 2.1 U/mL in the reusable bioreactor. After six perfusion harvests in the single-use, packed-bed bioreactor, the combined ALKP production was 16.2 U/mL compared to 17.4 U/mL in the reusable bioreactor in batch mode. Multiple batch culture runs in the pitched-blade bioreactor would be required to match the output of a single run in the packed-bed bioreactor in perfusion mode. Results demonstrate that there are no significant differences between the reusable and single-use systems for bench-scale production of recombinant proteins. Our results also suggest that the CelliGen BLU packed-bed bioreactor, when operated in perfusion mode, is superior to the CelliGen BLU pitched-blade bioreactor when operated in batch mode, confirming our studies from 2012...

Citation:
Hatton T, Barnett S, Sha M, Rashid K. A Comparative Bioreactor Vessel Study: Conventional Reusable Glass and Single-Use Disposables for the Production of Alkaline Phosphatase. BioProcess J, 2013; 12(1): 21-28. dx.doi.org/10.12665/J121.Rashid.

Posted online May 30, 2013.

 
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