Development and monitoring of an integrated bioprocess for production of a potential malaria vaccine with Pichia pastoris

An engineered variant of the yeast Pichia pastoris was used to produce the potential malaria vaccine D1M1. This paper describes the implementation of an expanded bed adsorption chromatography step into a sequential cultivation process of the yeast in order to link protein expression, cell release, and product capture of the secreted protein in a fully automated plant. The process is operated with a semi-continuous strategy for an integrated production. Process improvements have been achieved using design of experiments. Resulting purities of product achieved up to 87% with recoveries of 51% in a single downstream operation. Furthermore, a multivariate analysis of historical batch data was used to develop a golden batch model for process monitoring.     

超高效液相色谱串联质谱法测定烟草中的噁霉灵残留

为降低烟草中噁霉灵检测中的基质干扰，提高检测灵敏度和和稳定性，通过试验建立了以QuEChERS为样品前处理方法、液相色谱串联质谱仪为检测仪器的烟草中噁霉灵残留检测方法。样品加水浸润后，经丙酮提取、盐析试剂包除水、石墨化炭黑除杂后，采用Agilent ZORBAX 300SB-C8液相色谱柱（2.1 mm×100 mm，1.8 μm）分离，甲醇-水程序洗脱，ESI雾化，串联质谱MRM检测。结果发现，该方法在0.0025~0.0500 mg/L范围内线性良好，决定系数r²=0.9996，定量检测限1.06 μg/L，回收率在90.8%~92.0%之间，相对标准偏差为5.1%。运用该方法测定烟草中的噁霉灵，操作简单，检测精度高，满足CORESTA对烟草农残检测的要求。     

抗乙肝病毒表面抗原PreS1(20-47)的单链抗体在转基因烟草根分泌物中的初步表达

为探索利用植物根分泌表达重组蛋白的可行性，构建了含有抗乙肝病毒表面抗原PreS1(20—47)单链抗体(ScFv)基因的表达载体。该ScFv基因转化烟草后在烟草根部细胞的细胞质和内质网中获得表达。实验结果表明，5’端融合ER导向信号肽的重组ScFv可通过根分泌表达。     

腐殖酸对烤烟根系生长和生理活性的影响

对腐殖酸和基肥混施进行了烤烟盆栽试验。结果表明 ,腐殖酸能有效刺激根系的生理活性 ,提高根系抗坏血酸氧化酶、多酚氧化酶等呼吸酶的活性 ,增强根系的活力 ,并能有效促进烤烟根系生长 ,根干重、体积和长度均有提高。     

从烟草悬浮细胞中分离液泡前体的方法研究

通过跟踪含有液泡前体特异标记物的组分, 利用细胞壁酶水解胞壁制造原生质体, 并用真针头挤压的方法裂解细胞, 用蔗糖密度梯度离心的方法分离液泡前体. 通过相差显微镜观察, 发现所分离的液泡前体保持完整的结构. 利用不同细胞器的标记物抗体进行免疫标记检测, 证明所纯化的液泡前体不含其他细胞器, 具有较高的纯度.     

Indoor Air Quality: Exploring Policy Options to Reduce Human Exposures

Deciding between the different policy approaches available for reducing human exposures to indoor pollutants is an exceptionally complex task. These options can range from waiting until more definitive information is available to enacting regulatory standards, with many variations in between. This paper presents some of the factors policy-makers must consider in establishing indoor air quality policies, and the role researchers should play in ensuring that indoor air policies are based on the best available scientific information.     

On-line H NMR spectroscopic investigation of hydrogen bonding in supercritical and near critical CO2–methanol up to 35 MPa and 403 K

On-line NMR spectroscopy can beneficially be applied to studies of supercritical and near-critical fluids as an alternative to optical spectroscopy. Up to now high pressure NMR experiments are predominantly accomplished using custom made NMR batch reactors. The authors present a novel high pressure cell with displacement plunger for on-line NMR experiments on compressible fluids which can be used in conjunction with commercially available SCF NMR flow probes. The on-line technique offers advantages compared to stopped flow techniques such as enhanced control of mixture composition and reaction parameters as well as the facility of engagement into the reaction. The new apparatus is used for NMR studies on hydrogen bonding of methanol in near critical and supercritical carbon dioxide up to 403 K and 35 MPa for which data on the chemical shift of the hydroxyl group and methyl group are reported and interpreted.     

二氧化碳与磷化氢混合熏蒸防治储烟害虫试验

目前,储烟害虫防治普遍采用磷化铝片剂常规熏蒸法,磷化铝施用量为6～9g/m3,安全上存在一定的隐患。为了减少磷化铝用药量,于2003年应用微电脑控制的仓外混合熏蒸机,施用钢瓶装液化二氧化碳和56%磷化铝片剂,进行了二氧化碳与磷化氢混合熏蒸杀虫试验,以8g/m3磷化铝常规熏蒸法为对照。试验结果表明,磷化铝施用量为2.3g/m3、二氧化碳施用量为12g/m3,混合熏蒸结果达到100%杀死害虫。与对照相比,杀虫效果相同,混合熏蒸法具有磷化铝施用量较小、较为安全和有利于减少污染等优点。     

烟机异型件CAD/CAM系统的开发与应用

结合企业异型件CAD/CAM系统的开发与应用实际，简要说明了应用CAD/CAM技术是实现烟草机械异型件国产化的必要途径，论述了CAD/CAM系统的开发所依据的技术原理和涉及到的关键技术问题，分析了应用该系统所产生的社会经济效益．