Biopharmaceuticals development

Stephen Tuck is Vice President of Biopharmaceutical Development at Dynavax Technologies in Berkeley, California. He has over 14 years of experience in pharmaceutical chemistry. He was involved in the development of Fluad adju-vated flu vaccine as well as various subunit vaccines, adjuvants, vaccine conjugates, and protein therapeutics. He earned his B.Sc. and Ph.D. degrees from Imperial College, University of London, United Kingdom. 

The Biop harm Guide to Biopharmaceutical Development, A supplement to Bio-Pharm, Patrick Clinton, editor-in-chief, 2002. 

If analytical methods are at the heart of biopharmaceutical development and manufacturing, then protein concentration methods are the workhorse assays. A time and motion study of the discovery, development, and manufacture of a protein-based product would probably confirm the most frequently performed assay to be protein concentration. In the 1940s Oliver H. Lowry developed the Lowry method while attempting to detect miniscule amounts of substances in blood. In 1951 his method was published in the Journal of Biological Chemistry. In 1996 the Institute for Scientific Information (ISI) reported that this article had been cited almost a quarter of a million times, making it the most cited research article in history. This statistic reveals the ubiquity of protein measurement assays and the resilience of an assay developed over 60 years ago. The Lowry method remains one of the most popular colorimetric protein assays in biopharmaceutical development, although many alternative assays now exist. 

Use of Reversed-Phase Liquid Chromatography in Biopharmaceutical Development... 

The choice of HPLC as a separation technique in biopharmaceutical development may depend upon its scalability, even if its resolving power is less than other techniques. For example, HPLC may be used in preference to gel electrophoresis or capillary electrophoresis, where scaleup is difficult or impossible. 

The MS techniques described previously for characterization of the final recombinant protein product can be applied at all stages during process development. MS might be used upstream to define clone selection, processing format, and purification steps, and downstream to characterize the final product, ascertain lotto-lot reproducibility, determine stability, and define the formulation of biopharmaceutical molecules. Presented here are some examples found either in the literature or from our own experience in which MS has been found to be a useful or necessary tool. Potential limitations of MS methods are discussed, and when appropriate, other analytical methods are mentioned that can be alternatives to MS and are also efficient tools for biopharmaceutical development. 

The first step in biopharmaceutical development is the selection of a clone in a specific cell line. Whole-mass analysis, if possible, is a fairly simple and powerful tool at this stage to verify the successful expression and translation of the desired protein. VanAdrichem et al.65 described the use of MALDI MS to monitor protein expression in several mammalian cell lines like CHO DXB11, CHO SSF3, and hybridomas. Quantitative MALDI-TOF MS measurements of an IgG antibody and insulin during large-scale production in hybridoma cells were comparable to affinity chromatography results. 

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