(Dt,C) Optimal run orders.

Cost considerations have rarely been taken into account in optimum design theory. A few authors consider measurement costs, i.e. the costs associated with a particular factor level combination. A second cost approach results from the fact that it is often expensive to change factor levels from one observation to another. We refer to these costs as transition costs. In view of cost minimization, one should minimize the number of factor level changes. However, there is a substantial likelihood that there is some time order dependence in the results. Consequently, when considering both time order dependence and transition costs, an optimal ordering is not easy to find. There is precious little in the literature on how to select good time order sequences for arbitrary design problems and up to now, no thorough analysis of both costs is found in the literature. For arbitrary design problems, our proposed design algorithm incorporates cost considerations in optimum design construction and enables one to compute cost-efficient run orders that are optimally balanced for time trends. The results show that cost considerations in the construction of trend-resistant run orders entail considerable reductions in the total cost of an experiment and imply a large increase in the amount of information per unit cost.Optimal; Run orders;

Joint Identification of Location and Dispersion Effects in Unreplicated Two-Level Factorials

Most procedures that have been proposed to identify dispersion effects in unreplicated factorial designs assume that location effects have been identified correctly. Incorrect identi- fication of location effects may impair subsequent identification of dispersion effects. We develop a model for joint identification of location and dispersion effects that can reliably identify active effects of both types. The joint model is estimated using maximum likelihood, and hence effect selection is done using a specially derived information criterion. An exhaustive search through a limited version of the space of possible models is conducted. Both a single-model output and model averaging are considered. The method is shown to be capable of identifying sensible location-dispersion models that are missed by methods that rely on sequential estimation of location and dispersion effects

Statistical techniques for quality improvement

Call number: LD2668 .R4 STAT 1989 G37Master of ScienceStatistic

Development of a new medium containing date syrup for production of bleomycin by Streptomyces mobaraensis ATCC 15003 using response surface methodology

A combined statistical approach of orthogonal design and polynomial regression were applied to optimize the composition and concentration of a liquid fermentation medium for the production of bleomycin (BLM) by Streptomyces mobaraensis. Optimal conditions for maximal productivity were determined based on eight parameters at three different levels. The sources of carbon and nitrogen concentration and their interactions with other precursors were found to be statistically significant factors. When date syrup was used as an additional carbon source, higher BLM amount was obtained in comparison to glucose. It was found that the optimum nitrogen source was achieved with the use of soyabean meal. The combined orthogonal design and response surface methodology predicted optimal conditions for production of BLM to be 138 mg dl-1. A confirmatory experiment of the optimal medium composition produced 142 mg dl-1 in the fifth day fermentation at 30°C. The complex medium containing 40 gml-1 date syrup as additional carbon source enhanced the production of BLM by 73%. The combined statistical approach enabled rapid identification and integration of key medium parameters for optimizing secondary metabolite production and could be very useful in pharma-ceutical screening programs.Keywords: Bleomycin, Streptomyces mobaraensis, orthogonal design, medium optimization, date syrupAfrican Journal of Biotechnology Vol. 9(33), pp. 5450-5459, 16 August, 201

Biossíntese da proteína Catecol-O-metiltransferase membranar humana: optimização com recurso a desenho experimental Plackett-Burman e composto central

Catechol-O-methyltransferase (COMT, EC 2.1.1.6) is an S-adenosyl-L-methionine-dependent methyltransferase enzyme that catalyzes the methylation of catechol substrates (catecholamines, catecholestrogens). Physiologically, it is responsible for the elimination of biologically active or toxic catechols, making it a protein with great clinical relevance as therapeutic target in serious disorders, like schizophrenia and Parkinson´s disease. To fulfill pharmaceuticals requirements, new strategies of optimization and large-scale production of COMT enzyme are crucial. Statistical optimization approaches have demonstrated their enormous value in laboratory and industrial scale, namely in biotechnological production processes, in which an incremental enhancement can be a perpetual improvement. In this work, we aimed the optimization of recombinant human membrane-bound COMT (hMBCOMT) enzymatic activity yields following a statistical optimization as a solving approach. Plackett-Burman design was used as a first optimization step to identify which factors have a significant effect in hMBCOMT productivity and activity levels, and Response Surface Methodology (RSM), by a Central Composite Design (CCD), to optimize the process. We applied Brevibacillus choshinensis cells for the biosynthesis of hMBCOMT and a semi-defined medium for cell growth. This medium was subjected to a first screening using the Plackett–Burman design to evaluate the influence of the culture parameters (chemicals and physicals) in hMBCOMT enzymatic activity levels. Enzymatic activity were measured in a high performance liquid chromatography (HPLC) coupled to a coulochemical detector. Among the eleven variables tested, polypeptone, ammonium sulfate, glucose and temperature were selected owing to their significant effect on human MBCOMT enzymatic activity. The biological human MBCOMT activity obtained with the semi-defined medium in Plackett-Burman design were very promising, while were higher than the obtained with 2SYNm medium, a traditional growth medium for Brevibacillus cells of this work. Typically, we obtained values of 93nmol/h for hMBCOMT total enzymatic activity and 30 nmol/h/mg of specific activity with protein in its native form, without the use of any kind of detergents on protein solubilization step. Based on the results of Plackett–Burman design, a CCD was adopted to define optimal components concentration and temperature in order to maximize our response. The CCD model presented a multiple correlation coefficient value of 0.635 and a significant lack of fit, showing the lack aptness of the model to the process optimization and the failure to attain the optimal concentration of each variable.Catecol-O-metiltransferase (COMT, CE 2.1.1.6) é uma enzima metiltransferase dependente de S-adenosil-L-metionina (SAM) que catalisa a metilação de substratos catecóis (catecolaminas, catecolestrogénios). Fisiologicamente, é responsável pela eliminação de catecóis biologicamente activos ou tóxicos, tornando-a uma proteína de elevado interesse clínico e utilizada como alvo terapêutico em doenças graves, como a esquizofrenia e a doença de Parkinson. Para suprir as necessidades farmacêuticas, novas estratégias de otimização e produção em larga escala desta enzima são fundamentais. Abordagens de otimização estatística têm demonstrado o seu enorme valor à escala laboratorial e industrial, nomeadamente nos processos de produção biotecnológicos, em que um pequeno detalhe melhorado pode significar um grande passo para o sucesso. Neste trabalho, objetivou-se a otimização do nível de atividade enzimática da proteína recombinante COMT, na sua forma membranar, através do recurso a modelos de otimização estatística como uma abordagem resolutiva. Numa primeira fase de otimização e de seleção dos fatores mais significativos para a atividade enzimática da proteína em estudo foi utilizada a técnica de desenho experimental Plackett-Burman. Após esta seleção foi aplicada a Metodologia de Superfície de Resposta (RSM), através de desenho composto central (DCC), para otimização da concentração dos fatores que revelaram ser mais significativos e, consequentemente, do processo. Foi utilizado o sistema de expressão Brevibacillus choshinensis para a biossíntese da proteína membranar COMT e um meio semi-definido para o seu crescimento. Este meio foi submetido a uma primeira triagem através do desenho experimental Plackett-Burman, avaliando-se desta forma a influência dos parâmetros de cultura (produtos químicos e físicos) nos níveis de actividade enzimática da COMT membranar. Os níveis de actividade enzimática foram medidos num sistema de cromatografia líquida de alta eficiência acoplado a um detector amperométrico. Entre as onze variáveis testadas, a polipeptona, sulfato de amónio, glucose e temperatura foram as variáveis selecionadas dado o seu significativo efeito na actividade enzimática da COMT membranar. Os níveis de atividade enzimática obtidos nesta primeira triagem revelaram-se bastante promissores, sendo mais elevados do que os obtidos com o meio 2SYNm, meio de crescimento mais comum para as células usadas neste trabalho. Foram obtidos valores de 93nmol/h para a actividade enzimática total e cerca 30 nmol/h/mg de actividade enzimática específica com a proteína na sua forma nativa, sem o uso de qualquer tipo de detergentes no processo de solubilização. Com base nos resultados do desenho Plackett Burman foi aplicado o desenho Composto Central para a otimização dos quatro fatores em causa a fim de maximizar a nossa resposta. Este apresentou um valor do coeficiente de correlação múltipla de 0,635 e uma falta de ajuste significativa, demonstrando a falta de adequação do modelo para a otimização do processo