Polysorbate-80, or polysorbitan mono-oleate, a non-ionic surfactant commercially referred to as Tween-80, is commonly employed as an excipient in the biopharmaceutical industry due to its low toxicity as well as its ability to solubilize hydrophobic molecules. Thus, polysorbate-80 is employed as a stabilizer in order to control the formation of protein aggregates and larger particulates in biopharmaceutical drug substance and drug product formulations. Guidelines published by The International Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use (ICH) and the United States Pharmacopeia (USP) classify excipients such as polysorbate-80 as major components in biopharmaceutical formulations and specify that these types of compounds be quantified using a validated analytical method…
BioProcessing Journal Posts
Since the first gene therapy trials were conducted 25 years ago, there have been high expectations from the public, and much attention from investors, that previously incurable diseases would be cured by gene therapy. Still, despite numerous gene therapy clinical trials for many different indications, there are no approved gene therapy drugs in the United States. In 1999, one gene therapy patient died during clinical trials, the first ever. This highly publicized event led to heightened regulatory scrutiny over all such trials. Then in 2003 and 2005, three subjects developed leukemia as a direct consequence of gene therapy; one of them eventually passed away. The regulatory response stemming from these incidents led to greater regulatory oversight in gene therapy, as compared to other investigational drugs and biologics…
Optimal process development creates unit operations that effectively generate, separate, and concentrate a broad array of products. Historically, tangential flow filtration (TFF) process capabilities have been limited by technological and flow restrictions. Recent innovations in TFF module design have dramatically increased the capabilities of TFF to better achieve processing objectives. NCSRT has established a best practices protocol for developing clarification, fractionation, and concentration processes for mammalian, bacteria, yeast, insect, and virus based production systems. This article presents the development platform, supplemented with application-specific expertise…
Baculoviruses have found many uses in the field of biology, including as a control strategy for major insect pests such as the gypsy moth, the corn earworm, and the velvetbean caterpillar, for high level expression of recombinant proteins in insect cells, and, more recently, as gene delivery vehicles into mammalian cells. Some of these uses entail the need for isolation of baculoviral DNA for molecular biology studies. Currently, there are three ways in which baculoviral DNA can be obtained: 1) from the infection of insect larvae or cell cultures and the recovery of occlusion bodies (OB) containing viral particles, from which the DNA is then extracted; 2) from the recovery of extracellular virus (ECV) or budded virus (BV), usually from infected cell cultures; or 3) from total intracellular DNA harvested from infected cell cultures early in the infection cycle…
The developing biotechnology community may offer solutions and hope for recent world events that have focused attention on the vulnerability of the world’s population. Concerns about new pandemics have been raised by the emergence of new influenza strains and the re-emergence of older and even more highly virulent strains. In addition, there are fears that bioterrorism could involve agents such as anthrax or smallpox, and these threats become even more of a concern when you consider the increased mobility of such organisms via today’s commercial aviation. The ability of the biomedical community to respond rapidly to these shifting threats is more important than ever…
One of the major aims of modern biotechnology companies that are producing recombinant therapeutic proteins is to focus on timeline reduction of critical cell line selection and process optimisation studies in order to minimise the time and financial constraints of early development products. This “minimalist paradigm” of maximising early development throughput with minimal capital/operational outlays is a key driver for implementation of novel analytical technologies which can be applied at-line to process instrumentation. The large-scale production of recombinant therapeutics in the biopharmaceutical industry relies on in-process monitoring of product titre. Traditional titre determination methods, including enzyme-linked immunosorbent assay (ELISA) and protein A high performance liquid (immunoaffinity) chromatography (Protein A HPLC), are time consuming, and often reliant on analytical support from separate specialist teams/departments requiring detailed scientific knowledge and extensive training, with expensive capital outlay utilising large equipment…
The significance of tissue culture is that an understanding of animal physiology will come from a study of the basic elements of the animal (the cell) in isolated, pure form (clonal culture), under defined conditions (hormonally defined media). This should be the fundamental approach to animal physiology for the foreseeable future. The problem with this formulation in 1958, when I began work on tissue culture, was that cells in culture did not display the differentiated properties of the tissue of origin…
For more than a decade, transgenic plants have been investigated as alternatives to microbial, mammalian cell, and transgenic animal systems for recombinant protein production. The main advantages of using plants as “bioreactors” are that the cost of upstream production (i.e. biomass creation) is low; plants do not carry viruses and other pathogens dangerous to humans such as human immunodeficiency virus (HIV), prions, hepatitis viruses and so on; and as eukaryotes, plants are capable of producing bioactive proteins. Numerous recombinant proteins have been expressed in various plant hosts, and some recombinant proteins are in various stages of clinical trials…
The baculovirus expression vector system, which is based on infecting insect cells with recombinant Autographa californica nuclear polyhedrosis virus (AcNPV), is one of the most commonly used eukaryotic expression systems aimed at producing functionally active mammalian proteins. It offers advantages such as high-level protein expression and post-translational processing capabilities that are extremely important to the biological activity of certain proteins. This system utilizes a strong promoter of the very late gene, polyhedrin, to drive heterologous protein overexpression. Nevertheless, in order to generate milligram amounts of recombinant proteins, cell culture often needs to be scaled up to as much as 25 liters….
The outstanding success and safety record of first generation monoclonal products has created an immense increase in the number of product candidates that need to be evaluated clinically. The concept of platform purification has emerged in response to this need. A platform is a semigeneric, multistep purification procedure that can be applied to a wide range of monoclonal antibodies without extensive method-scouting and optimization. This approach can substantially accelerate process development and hasten inception of clinical trials…