Process Economics of Animal Cell and Bacterial Fermentations: A Case Study Analysis of Tissue Plasminogen Activator

Datar, Rajiv; Cartwright, Terence; Rosén, Carl-Gustaf

doi:10.1038/nbt0393-349

Cited by 170 publications

(89 citation statements)

References 12 publications

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“…The recombinant product produced by a mammalian fermentation system was harvested from the culture medium. Attracted by simplicity and economy of production, investigators made a number of efforts in producing rtPA from bacteria, especially from Escherichia coli (10,13,30). Numerous strategies have been proposed to overcome the problems of low yield and the formation of inclusion bodies, which result in misfolding and in an inactive enzyme.…”

mentioning

confidence: 99%

Secretion of Active Recombinant Human Tissue Plasminogen Activator Derivatives in Escherichia coli

Manosroi¹,

Tayapiwatana

Götz

et al. 2001

Appl Environ Microbiol

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The DNA fragment coding for kringle 2 plus serine protease domains (K2S) of tissue plasminogen activator (tPA) was inserted into a phagemid vector, pComb3HSS. In the recombinant vector, pComb3H-K2S, the K2S gene was fused to gpIII of ⌽M13 and linked to the OmpA signal sequence. The resulting gene, rK2S-gpIII, was inducibly expressed in Escherichia coli XL-1 Blue. The protein was presented on the phage particle. To stop the expression of gpIII, a stop codon between K2S and the gpIII gene was inserted by site-directed mutagenesis. This mutated vector, MpComb3H-K2S, was transformed in XL-1 Blue. After induction with IPTG (isopropyl-␤-Dthiogalactopyranoside), rK2S was found both in the periplasm as an inactive form of approximately 32% and in the culture supernatant as an active form of approximately 68%. The secreted form of rK2S was partially purified by ammonium sulfate (55%) precipitation. The periplasmic form was isolated from whole cells by chloroform extraction. The fibrin binding site of kringle 2 was demonstrated in all expressed versions (phage-bound, periplasmic, and secreted forms) using the monoclonal anti-kringle 2 antibody (16/B). Only the secreted form of rK2S revealed a fibrinogen-dependent amidolytic activity with the specific activity of 236 IU/g. No amidolytic activity of rK2S was observed in either the periplasmic or the phage-bound form. The secretion of rK2S as an active enzyme offers a novel approach for the production of the active-domain deletion mutant tPA, rK2S, without any requirements for bacterial compartment preparation and in vitro refolding processes. This finding is an important technological advance in the development of large-scale, bacteriumbased tPA production systems.

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confidence: 99%

Secretion of Active Recombinant Human Tissue Plasminogen Activator Derivatives in Escherichia coli

Manosroi¹,

Tayapiwatana

Götz

et al. 2001

Appl Environ Microbiol

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“…Datar, Cartwright, and Rosen (1993) illustrated how the expression system could have a major impact on the total number of required processing steps and hence the economic viability of a product. They compared the costs of producing the recombinant protein, rtPA, in E. coli and CHO cells.…”

Section: Cost Modellingmentioning

confidence: 99%

Modelling biopharmaceutical manufacture: Design and implementation of SimBiopharma

Farid

Washbrook

Titchener‐Hooker

2007

Computers & Chemical Engineering

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This paper presents the implementation of a conceptual framework, for modelling a biopharmaceutical manufacturing plant, into a prototype decision-support tool, SimBiopharma. The tool's scope covers the ability to evaluate manufacturing alternatives in terms of cost, time, yield, resource utilisation and risk. Incorporating uncertainty means that investment appraisal can be based on both the expected outputs and the likelihood of achieving them. A hierarchical approach to represent the key activities in a manufacturing process is introduced. Emphasis is placed on how a closer integration of bioprocess and business process modelling can be achieved by capturing common information in an object-oriented environment, G2 (Gensym Corporation, Cambridge, MA). The key features of SimBiopharma are highlighted; these include interactive graphics, task-oriented representation and dynamic simulation which create a much more flexible environment for modelling processes. Examples of typical outputs generated by SimBiopharma, when addressing the impact of manufacturing options on strategic operational and financial indicators, are given.

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“…Large-scale processes for production of recombinant proteins from E. coli have been reported (Strandberg et al, 1991), even though, in certain expression strategies, the recombinant protein is expressed in an insoluble form. Nevertheless, bacterial expression may still be less economical (Datar et al, 1993), in light of the rapid progress in biotechnology.…”

Section: K S S S T a Y M Q L S S L T S E D S A V Y Y Y Cmentioning

confidence: 99%

Passive Immunization Against Dental Caries and Periodontal Disease: Development of Recombinant and Human Monoclonal Antibodies

Abiko

2000

Critical Reviews in Oral Biology & Medicine

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Indigenous micro-organisms in the oral cavity can cause two major diseases, dental caries and periodontal diseases. There is neither agreement nor consensus as to the actual mechanisms of pathogenesis of the specific virulence factors of these micro-organisms. The complexity of the bacterial community in dental plaque has made it difficult for the single bacterial agent of dental caries to be determined. However, there is considerable evidence that Streptococcus mutans is implicated as the primary causative organism of dental caries, and the cell-surface protein antigen (SA I/Il) as well as glucosyltransferases (GTFs) produced by S. mutans appear to be major colonization factors. Various forms of periodontal diseases are closely associated with specific subgingival bacteria. Porphyromonas gingivalis has been implicated as an important etiological agent of adult periodontitis. Adherence of bacteria to host tissues is a prerequisite for colonization and one of the important steps in the disease process. Bacterial coaggregation factors and hemagglutinins likely play major roles in colonization in the subgingival area. Emerging evidence suggests that inhibition of these virulence factors may protect the host against caries and periodontal disease. Active and passive immunization approaches have been developed for immunotherapy of these diseases. Recent advances in mucosal immunology and the introduction of novel strategies for inducing mucosal immune responses now raise the possibility that effective and safe vaccines can be constructed. In this regard, some successful results have been reported in animal experimental models. Nevertheless, since the public at large might be skeptical about the seriousness of oral diseases, immunotherapy must be carried out with absolute safety. For this goal to be achieved, the development of safe antibodies for passive immunization is significant and important. In this review, salient advances in passive immunization against caries and periodontal diseases are summarized, and the biotechnological approaches for developing recombinant and human-type antibodies are introduced. Furthermore, our own attempts to construct single-chain variable fragments (ScFv) and human-type antibodies capable of neutralizing virulence factors are discussed.

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Process Economics of Animal Cell and Bacterial Fermentations: A Case Study Analysis of Tissue Plasminogen Activator

Cited by 170 publications

References 12 publications

Secretion of Active Recombinant Human Tissue Plasminogen Activator Derivatives in Escherichia coli

Secretion of Active Recombinant Human Tissue Plasminogen Activator Derivatives in Escherichia coli

Modelling biopharmaceutical manufacture: Design and implementation of SimBiopharma

Passive Immunization Against Dental Caries and Periodontal Disease: Development of Recombinant and Human Monoclonal Antibodies

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