Regulation of advanced therapy medicinal products in Europe and the role of academia

Background aimsAdvanced therapy medicinal products (ATMP) are gene therapy, somatic cell therapy or tissue-engineered products regulated under (EC) No. 1394/2007 to ensure their free movement within the European Union while guaranteeing the highest level of health protection for patients. Academic good manufacturing practice (GMP) centers are major contributors in the development of ATMPs and this study assessed the impact of regulations on them.MethodsEuropean academic and non-industrial facilities (n = 747) were contacted, and a representative sample of 50 replied to a detailed questionnaire. Experienced centres were further selected in every Member State (MS) for semi-structured interviews. Indicators of ATMP production and development success were statistically assessed, and opinions about directive implementation were documented.ResultsFacilities experienced in manufacturing cell therapy transplant products are the most successful in developing ATMPs. New centres lacking this background struggle to enter the field, and there remains a shortage of facilities in academia participating in translational research. This is compounded by heterogeneous implementation of the regulations across MS.ConclusionsGMP facilities successfully developing ATMPs are present in all MS. However, the implementation of regulations is heterogeneous between MS, with substantial differences in the definition of ATMPs and in the approved manufacturing environment. The cost of GMP compliance is underestimated by research funding bodies. This is detrimental to development of new ATMPs and commercialization of any that are successful in early clinical trials. Academic GMP practitioners should strengthen their political visibility and contribute to the development of functional and effective European Union legislation in this field.     

Decisional tool to assess current and future process robustness in an antibody purification facility

Increases in cell culture titers in existing facilities have prompted efforts to identify strategies that alleviate purification bottlenecks while controlling costs. This article describes the application of a database‐driven dynamic simulation tool to identify optimal purification sizing strategies and visualize their robustness to future titer increases. The tool harnessed the benefits of MySQL to capture the process, business, and risk features of multiple purification options and better manage the large datasets required for uncertainty analysis and optimization. The database was linked to a discrete‐event simulation engine so as to model the dynamic features of biopharmaceutical manufacture and impact of resource constraints. For a given titer, the tool performed brute force optimization so as to identify optimal purification sizing strategies that minimized the batch material cost while maintaining the schedule. The tool was applied to industrial case studies based on a platform monoclonal antibody purification process in a multisuite clinical scale manufacturing facility. The case studies assessed the robustness of optimal strategies to batch‐to‐batch titer variability and extended this to assess the long‐term fit of the platform process as titers increase from 1 to 10 g/L, given a range of equipment sizes available to enable scale intensification efforts. Novel visualization plots consisting of multiple Pareto frontiers with tie‐lines connecting the position of optimal configurations over a given titer range were constructed. These enabled rapid identification of robust purification configurations given titer fluctuations and the facility limit that the purification suites could handle in terms of the maximum titer and hence harvest load. © 2012 American Institute of Chemical Engineers Biotechnol. Prog., 28: 1019–1028, 2012     

Cronobacter species (formerly known as Enterobacter sakazakii) in powdered infant formula: a review of our current understanding of the biology of this bacterium

Cronobacter species (formerly known as Enterobacter sakazakii) are opportunistic pathogens that can cause necrotizing enterocolitis, bacteraemia and meningitis, predominantly in neonates. Infection in these vulnerable infants has been linked to the consumption of contaminated powdered infant formula (PIF). Considerable research has been undertaken on this organism in the past number of years which has enhanced our understanding of this neonatal pathogen leading to improvements in its control within the PIF production environment. The taxonomy of the organism resulted in the recognition of a new genus, Cronobacter, which consists of seven species. This paper presents an up-to-date review of our current knowledge of Cronobacter species. Taxonomy, genome sequencing, current detection protocols and epidemiology are all discussed. In addition, consideration is given to the control of this organism in the manufacturing environment, as a first step towards reducing the occurrence of this pathogen in PIF.     

Determination and control of low-level amino acid misincorporation in human thioredoxin protein produced in a recombinant Escherichia coli production system

Escherichia coli is used extensively in the production of proteins within biotechnology for a number of therapeutic applications. Here, we discuss the production and overexpression of the potential biopharmaceutical human thioredoxin protein (rhTRX) within E. coli. Overexpression of foreign molecules within the cell can put an enormous amount of stress on the translation machinery. This can lead to a misfiring in the construction of a protein resulting in populations differing slightly in amino acid composition. Whilst this may still result in a population of active molecules being expressed, it does present significant problems with molecules that are destined for clinical applications. Amino acid misincorporation of this subset could potentially result in antibodies being raised to these unnatural proteins. Cross-reaction with a patient's endogenous thioredoxin could then lead to an autoimmune phenomena and serious health implications. Generally, the issue of misincorporation appears not to be a routine regulatory concern (see ICH Q6B guidelines). Therefore, amino acid misincorporation may not have been detected, much less explored in the clinic as the occurrence or absence of these random errors is not routinely reported. Using current technologies based on proteomics, the ability to find misincorporation critically depends upon the criteria for matching theoretical and experimental mass spectrometry data. Additionally, isolation and extraction of these mistranslated proteins from the production process is both difficult and expensive. Therefore, it is advantageous to find routes for removing their production during the upstream phase. In this study, we show how modern proteomic technology can be used to identify and quantify amino acid misincorporation. Using these techniques we have shown how manipulation of gene sequence and scoping of fermentation media composition can lead to the reduction and elimination of these misincorporations in rhTRX.     

生物有机无机复合肥的研制与效果研究

研制了一种融微生物肥有机肥和无机肥于一体的新型复合肥料。测定结果,含微生物活菌总量为2．5亿个／g,其中固氮、解磷、解钾功能做生物活菌量达1．0亿个／g,有机干物质34．3－37．3％,含氮（N）7.8－8．9％,全磷（P2O5）5．2－5．5％,全钾（K2O）4．8—5．0％。毒性试验结果表明,该肥无毒无害,对环境不构成危害。生产过程中采取了有机物料发酵和功能微生物大量繁殖一次同步完成,添加除臭微生物、机械自动翻拌等工艺。田间试验表明,与等养分或等价格投入的化肥相比,增产9－18％,对多种经济作物品质有明显的改善作用,且有一定的改土培肥效应。     

Development of a Device in Detection of Glaucoma for Rural Eye Care Using Additive Manufacturing and TRIZ

Purpose: The main purpose of this study is to develop a device for the indicative measurement of intraocular pressure (IOP) of eyeball, a key cause for glaucoma. In early diagnosis and treatment of glaucoma accurate measurement of IOP is important. The methods and devices which are available for the measurement of IOP have their own limitations which cause discomfort to the patients during measurement and needs anesthesia. There is a dare need of a device for the measurement of intraocular pressure by making the contact of plunger with closed eyelid eliminating the need of anesthesia and expert ophthalmologist. Method: Additive manufacturing (AM) is an era of technical development and innovation. Developing a device for detecting glaucoma by using AM and TRIZ ‘The theory of inventive problem solving’ (A Collaborative approach) can overcome the disadvantages that classic tonometer have. The field of Ophthalmology will be experiencing a paradigm shift towards the use of collaborative approach of TRIZ with AM. The developed new device was tested on 40 patient’s eye at Government Hospital Bhandara, (M. S.), India. The results of new device were cross verified by expert clinicians using calibrated Schiotz’s tonometer and digital palpation technique. Result: The developed new device was tested on patient’s eye through eyelid and results were compared with calibrated Schiotz’s tonometer. The results from the new device were found in good agreement with results from Schiotz’s tonometer with the average error of 0.033 ± 0.18 (mean ± SD) mm of Hg and mean relative error was -0.0018 ±0.0096 (mean ± SD). Conclusion: There is a substantial need for early detection and diagnosis of glaucoma in rural and remote areas (worldwide). A new device for detection of glaucoma using AM and TRIZ was introduced in this paper and measurements by the new device were by currently well accepted Schiotz’s tonometer. The new device will help the medical practitioners in rural and remote areas for early detection of glaucoma.     

经编人造血管顺应性的实验研究

对EP系列经编人造血管的径向顺应性和纵向顺应性进行了体外测试和研究,并分析了不同制备工艺对经编人造血管的径向顺应性的影响。实验结果表明,紧密化处理和波纹化热定性对人造血管的径向顺应性影响较大。测试压力段的选择对人造血管成品的纵向顺应性有较大的关系。     

Alkaline preserved herring by-products in feed for Atlantic salmon (Salmo salar L.)

The aim of the present study was to investigate the digestibility, growth and slaughter quality in Atlantic salmon (Salmo salar L.) fed a novel type of moist feed. The moist feed was prepared from alkaline preserved (pH 11.2) herring (Clupea harengus) filleting by-products, mixed with a crude binder based on seaweed before the feed was shaped in a pelleting process and the final structure was set immersing the pellets in an acidic bath. The feeding experiment was carried out with seawater-adapted salmon with an average initial weight of 1.7 kg. The moist feed and a control feed were fed to three groups of salmon with 190 fish per group. The coefficient of total tract apparent digestibility of fat, protein and energy in fish fed the moist feed was 0.96, 0.81 and 0.87, respectively. The weight gain for fish fed moist feed was significantly higher compared to salmon fed commercially extruded control feed. Carcass quality data showed that fish fed the moist feed had significantly higher dressing out percentage and less visceral fat compared to fish fed the control feed. It is concluded that alkaline preserved fish by-products can be efficiently utilized in a novel moist feed technology, yielding good growth and digestibility in Atlantic salmon.