Full Description
More then 20 years have passed now since the first recombinant protein producing microorganisms have been developed. In the meanwhile, numerous proteins have been produced in bacteria, yeasts and filamentous fungi, as weIl as higher eukaryotic cells, and even entire plants and animals. Many recombinant proteins are on the market today, and some of them reached substantial market volumes. On the first sight one would expect the technology - including the physiology of the host strains - to be optimised in detail after a 20 year's period of development. However, several constraints have limited the incentive for optimisation, especially in the pharmaceutical industry like the urge to proceed quickly or the requirement to define the production parameters for registration early in the development phase. The additional expenses for registration of a new production strain often prohibits a change to an optimised strain. A continuous optimisation of the entire production process is not feasible for the same reasons.
Contents
Cell-free translation systems.- Protein synthesis and co-translational folding in cell-free translation systems.- Metabolic burden and stress response.- The cellular response to unfolded proteins in the endoplasmic reticulum.- The effects of recombinant protein expression on the growth and metabolism of mammalian cells.- Mapping stresses in Escherichia coli to improve yield. Examining global gene regulation and « cell conditioning » strategies.- Cellular responses to strong overexpression of recombinant genes in Escherichia coli. DNA relaxation and cell death after induction of ?-glucosidase.- From Vitreoscilla hemoglobin (VHb) to a novel class of growth stimulating hemoglobin proteins.- Genetic stability and gene copy number effects.- Protein mass production in hybridomas and recombinant CHO cells.- Inducible gene copy number amplification for the production of heterologous proteins in Kluyveromyces lactis.- Antibiotic-free plasmid selection and maintenance in Bacteria.- Modelling of segregational plasmid instability of recombinant strain suspension of Escherichia coli.- Transcription, translation, and product formation in E. coli.- Production of optically pure aryl epoxides by recombinant E. coli carrying styrene monooxygenase. A new biocatalyst based on Pseudomonas fluorescens ST genes.- Translational problems associated with the rare arginine CGG in Escherichia coli. Frameshifhting at CGG codons.- Optimisation of the solubility of the recombinant Itk kinase domain in Escherichia coli.- Bacterial senescence and the oxidation paradox.- Metabolic approaches for the optimisation of recombinant fermentation processes.- Control and optimisation of cellular bottlenecks in recombinant protein production.- Expression and fermentation strategies for recombinant proteinproduction in Escherichia coli.- Transcription, translation, and product formation in microbial systems other than E. coli.- Overexpression of a Rhizopus oryzae lipase in Pichia pastoris strains containing multiple copies of the target gene.- Development of a heterologous gene expression system for use in Lactococcus lactis. A novel gram-positive expression system.- Metabolic network analysis for human therapeutic protein productions: effects of the P/O ratio.- Animal cell based expression systems — process optimisation.- Process-orientated metabolic engineering: cell lines with new properties in nutrient exploitation and protein glycosylation.- Influence of the metabolic status of packaging cells on retroviral vector production.- Optimizing the production of recombinant prion protein from CHO cells.- Recombinant protein production by transient transfection of suspension-growing cells.- Comparison of different microbial expression systems.- Production of recombinant human trypsinogen in Escherichia coli and Pichia pastoris. A comparison of expression systems.- Secretion of homologous and heterologous recombinant proteins in Escherichia coli and other gram-negative bacteria by using a new secretion system.- Monitoring of genes that respond to overproduction of insoluble recombinant proteins in Escherichia coli and Bacillus subtilis.- Model supported optimization of fed-batch fermentations for recombinant protein production.- Protease secretion capacity and performance analysis of recombinant Bacillus species.- Authors Index.
