Control of culture environment for improved polyethylenimine-mediated transient production of recombinant monoclonal antibodies by CHO cells

In this study we describe optimization of polyethylenimine (PEI)-mediated transient production of recombinant protein by CHO cells by facile manipulation of a chemically defined culture environment to limit accumulation of nonproductive cell biomass, increase the duration of recombinant protein production from transfected plasmid DNA, and increase cell-specific production. The optimal conditions for transient transfection of suspension-adapted CHO cells using branched, 25 kDa PEI as a gene delivery vehicle were experimentally determined by production of secreted alkaline phosphatase reporter in static cultures and recombinant IgG(4) monoclonal antibody (Mab) production in agitated shake flask cultures to be a DNA concentration of 1.25 mu g 10(6) cells(-1) mL(-1) at a PEI nitrogen: DNA phosphate ratio of 20:1. These conditions represented the optimal compromise between PEI cytotoxicity and product yield with most efficient recombinant DNA utilization. Separately, both addition of recombinant insulin-like growth factor (LR3-IGF) and a reduction in culture temperature to 32 degrees C were found to increase product titer 2- and 3-fold, respectively. However, mild hypothermia and LR3-IGF acted synergistically to increase product titer 11-fold. Although increased product titer in the presence of LR3-IGF alone was solely a consequence of increased culture duration, a reduction in culture temperature post-transfection increased both the integral of viable cell concentration (IVC) and cell-specific Mab production rate. For cultures maintained at 32 degrees C in the presence of LR3-IGF, IVC and qMab were increased 4- and 2.5-fold, respectively. To further increase product yield from transfected DNA, the duration of transgene expression in cell populations maintained at 32 C in the presence of LR3-IGF was doubled by periodic resuspension of transfected cells in fresh media, leading to a 3-fold increase in accumulated Mab titer from similar to 13 to similar to 39 mg L-1. Under these conditions, Mab glycosylation at Asn297 remained essentially constant and similar to that of the same Mab produced by stably transfected GS-CHO cells. From these data we suggest that the efficiency of transient production processes (protein output per rDNA input) can be significantly improved using a combination of mild hypothermia and growth factor(s) to yield an extended activated hypothermic synthesis.

Computer-assisted production scheduling, planning and control in foundries

The present study describes a pragmatic approach to the implementation of production planning and scheduling techniques in foundries of all types and looks at the use of `state-of-the-art' management control and information systems. Following a review of systems for the classification of manufacturing companies, a definitive statement is made which highlights the important differences between foundries (i.e. `component makers') and other manufacturing companies (i.e. `component buyers'). An investigation of the manual procedures which are used to plan and control the manufacture of components reveals the inherent problems facing foundry production management staff, which suggests the unsuitability of many manufacturing techniques which have been applied to general engineering companies. From the literature it was discovered that computer-assisted systems are required which are primarily `information-based' rather than `decision based', whilst the availability of low-cost computers and `packaged-software' has enabled foundries to `get their feet wet' without the financial penalties which characterized many of the early attempts at computer-assistance (i.e. pre-1980). Moreover, no evidence of a single methodology for foundry scheduling emerged from the review. A philosophy for the development of a CAPM system is presented, which details the essential information requirements and puts forward proposals for the subsequent interactions between types of information and the sub-system of CAPM which they support. The work developed was oriented specifically at the functions of production planning and scheduling and introduces the concept of `manual interaction' for effective scheduling. The techniques developed were designed to use the information which is readily available in foundries and were found to be practically successful following the implementation of the techniques into a wide variety of foundries. The limitations of the techniques developed are subsequently discussed within the wider issues which form a CAPM system, prior to a presentation of the conclusions which can be drawn from the study.

An approach to the development of expert systems within production planning and control

The thesis presents an account of an attempt to utilize expert systems within the domain of production planning and control. The use of expert systems was proposed due to the problematical nature of a particular function within British Steel Strip Products' Operations Department: the function of Order Allocation, allocating customer orders to a production week and site. Approaches to tackling problems within production planning and control are reviewed, as are the general capabilities of expert systems. The conclusions drawn are that the domain of production planning and control contains both `soft' and `hard' problems, and that while expert systems appear to be a useful technology for this domain, this usefulness has by no means yet been demonstrated. Also, it is argued that the main stream methodology for developing expert systems is unsuited for the domain. A problem-driven approach is developed and used to tackle the Order Allocation function. The resulting system, UAAMS, contained two expert components. One of these, the scheduling procedure was not fully implemented due to inadequate software. The second expert component, the product routing procedure, was untroubled by such difficulties, though it was unusable on its own; thus a second system was developed. This system, MICRO-X10, duplicated the function of X10, a complex database query routine used daily by Order Allocation. A prototype version of MICRO-X10 proved too slow to be useful but allowed implementation and maintenance issues to be analysed. In conclusion, the usefulness of the problem-driven approach to expert systems development within production planning and control is demonstrated but restrictions imposed by current expert system software are highlighted in that the abilities of such software to cope with `hard' scheduling constructs and also the slow processing speeds of such software can restrict the current usefulness of expert systems within production planning and control.

New measures of job control, cognitive demand and production responsibility

Recent research has highlighted several job characteristics salient to employee well-being and behavior for which there are no adequate generally applicable measures. These include timing and method control, monitoring and problem-solving demand, and production responsibility. In this article, an attempt to develop measures of these constructs provided encouraging results. Confirmatory factor analyses applied to data from 2 samples of shop-floor employees showed a consistent fit to a common 5-factor measurement model. Scales corresponding to each of the dimensions showed satisfactory internal and test–retest reliabilities. As expected, the scales also discriminated between employees in different jobs and employees working with contrasting technologies.

A production system and its control

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Control parameters in the batchwise production of synthetic resins

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Application of computer control to the production of synthetic resins

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Community control in cellular protein production : consequences for amino acid starvation

The authors thank M. C. Romano, I. Stansfield, L. Ciandrini, A. Kort, and M. Rehberg for helpful discussions. This work was funded by BBSRC grants BB/F00513/X1 and BB/G010722, and the Scottish Universities Life Science Alliance (SULSA).

Community control in cellular protein production : consequences for amino acid starvation

The authors thank M. C. Romano, I. Stansfield, L. Ciandrini, A. Kort, and M. Rehberg for helpful discussions. This work was funded by BBSRC grants BB/F00513/X1 and BB/G010722, and the Scottish Universities Life Science Alliance (SULSA).