Real time PCR - European Pharmaceutical Review

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PAT ISSUE 2009 Design theory Of the conceptual models that are used in design theory 16-18 much attention has been given to the Hubka-Eder (HE) model 19-20 due to its generality and applicability to any technical system for manufacturing. Most applications are found in mechanical and electrical engineering although the theoretical model is as applicable for chemical and biological products. The HE-model is built around the transformation process (TrP) and its applied technologies (Tg) to transform the inputs, being the sums of operands (ΣOd1), i.e. the raw materials, properties (ΣPr1) and secondary components (ΣSecIn), into outputs (ΣOd2, ΣPr2, and ΣSecOut) (see Figure 2). The model then describes and analyses all effects that are exerted on the TrP. That includes the technical systems involved (ΣTS), all information systems (ΣIS) and all management and goal-setting systems (ΣM&GS). Thus, processing machines such as blending vessels, freezers, and robots are example of technical units and systems ΣTS) which transform chemical or biological reactants and additives by causing reaction, interaction or separation but without being transformed themselves. Data collection systems such as data processing software and batch reporting procedures are included within the ΣIS while protocols, procedures and quality guidelines such as GMP and operational SOPs for the TrP act to manage to reach set manufacturing goals and quality criteria in the ΣM&GS. The purpose with the design model is to carry out a systematical investigation of effects in all steps involved. The interaction chart in Figure 2 (see page 33) illustrates the top level of approaching the system; a more detailed chart on a second or third level would elucidate in a much more thorough and informative way the multitude of interacting effects. This analysis should then lead to the identification of alternative, potentially better design solutions. By this systematic approach too early fixations with un-reflected solutions is avoided. The HE-model has recently been adapted by us for biotechnology products and systems by adding a separate biological systems entity, ΣBS, to the design concept with the objective of deepening the functional analysis of the cross-interactions between the participating subsystems 21 . By that, complex biotechnology processes such as cell culturing procedures that exploit the transformation capacity of the biological systems per se, are easier to describe and analyse thoroughly. As we have brought up in a separate paper 1 , the concepts of PAT and QbD benefit from being analysed within the HE-modeling, since PAT includes the technical systems which should according to the PATguidelines 3 be used to direct the design of the manufacturing process and the QbD principles should interconnect and affect the product and manufacturing quality through its early integration in the system. Figure 3 on page 33 illustrates how the embryonic stem cell transformation procedures in principle could be related in a HE model. Note
SUBSCRIBE ISSUE 2009 PAT especially the fundamentally different way of describing the steps of the procedures compared to a typical laboratory manual. Unlike a SOP the functions of the steps are separated and thoroughly investigated in terms of effects on the procedural steps. Note also the influence of the active environment. The hESC process is subject to longterm exposure of contaminating disturbances from the environment – not possible to foresee, but to counteract. The biological variation, which is so important to measure or detect is included in the active environment. The depiction here is also integrating PAT and QbD in the conceptual design, in accordance with the regulators’ mindset, i.e. to bring in the quality aspects directly into the design process and with the help of available powerful PAT methods and tools. It is our interpretation of PAT that the HE modeling methodology can be considered as one of the useful PAT tools. Manufacturing aspects on stem cell production Scaled-up manufacturing of hESC and hESC-derived products requires access to exclusive biological systems, perfectly controlled and contamination-free manufacturing facilities, traceability of raw materials, xeno-free media, specialised equipment for handling cell culture, a variety of analytical instrument and quality control devices and, most importantly, very skilled and well trained staff and operators. The equipment should preferably be as automated as possible and the manufacturing should be performed in a clean room surrounding to reduce the risk for contamination. Most of the manufacturing equipment is commercially available such as freezing equipment, robots, incubators, microscopes, analysers, autopipets, vessels and other culturing devices. Consumables, single-use plastics, reagents and media are Figure 4: The HE depiction of the hESC manufacturing process in Figure 3 is detailed with additional effecting units to illustrate how the HE analysis is carried on for further investigating the interactions. supplied by numerous companies. The components are mostly used based on standard operating procedures and validated GMP protocols for preparation and testing. The procedures and protocols developed during the research and development phases should be framed into a large scale and adapted as much as possible to automation. This requires extensive further validation and reconsideration of effects caused by the scaled-up manufacturing systems. For example, procedures that can be automated in cell culturing robotic systems are expansion and maintenance of multiple cell lines, various sub-culturing steps, expanding cell numbers through the seeding of a number of flasks, harvesting and plating of cells for assays and screening, incubation in different flasks and plate formats, cell counting and viability measurement, processing of multilayer flasks, and automated plating. Successful automation of such procedures is accomplished for example in the Automation Partnership systems 22,23 . Other important functions the scaled-up manufacturing system should include are: ■ ■ ■ ■ ■ ■ Quality control of cell banks Quality control of input materials (culture media, factors, consumables) Quality control methods for manufactured cells (monitoring biomarkers; Functional testing of final cells, genomic analyses, histological analyses, microscopy, flow cytometry methods) Cryopreservation procedures (deep freezers for preserving cell materials at various stages of the process) Isolation procedures (dissection in microscopy, enzyme treatments) Preservation/storage of the product and shipping procedures Transforming the stem technology into an efficient manufacturing system by using conceptual design The complexity of these technical procedures for stem cell production makes it obvious that the transformation process into an efficient bio-mechatronic manufacturing system is difficult, <strong>time</strong>-consuming and, in the end, very cost dependent. To exploit modern manufacturing system development methods may GO TO CONTENTS PAGE 35
Page 1 and 2: TECHNOLOGY / KNOWLEDGE / INNOVATION
Page 3 and 4: EDITORIAL BOARD Dr Anthony Davies H
Page 5 and 6: ISSUE ONE 2009 Contents Index to Ad
Page 7 and 8: © 2009 Thermo Fisher Scientific In
Page 9 and 10: SUBSCRIBE ISSUE 2009 qPCR Figure 4:
Page 11 and 12: SUBSCRIBE ISSUE 2009 qPCR chromosom
Page 13 and 14: SUBSCRIBE ISSUE 2009 qPCR assays. T
Page 15 and 16: Single Cell Gene Expression from Fl
Page 17 and 18: © 2008 Thermo Fisher Scientific In
Page 19 and 20: SUBSCRIBE ISSUE 2009 HIGH CONTENT S
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Page 23 and 24: More Imaging Speed Introducing the
Page 25 and 26: SUBSCRIBE ISSUE 2009 LAB AUTOMATION
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Page 29 and 30: SUBSCRIBE ISSUE 2009 THERMAL ANALYS
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Page 33: SUBSCRIBE ISSUE 2009 PAT a bioproce
Page 37 and 38: SUBSCRIBE ISSUE 2009 PAT extensive
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