Design of high productivity sequential multi-column chromatography for antibody capture

An integrated experimental and modeling approach for the design of sequential multi-column chromatography (SMCC) is presented to maximize productivity in bioprocessing. The approach consists of three steps: (1) single-column model development and validation, (2) multi-column model development and validation, and (3) productivity optimization. The integrated use of process experimentation and modeling enables sufficient process understanding to be gained during process development such that the optimal SMCC design is found even with limited time and materials. The application of the approach is demonstrated by determining the optimal SMCC design that maximizes the capture of human IgG by a silica-based protein A adsorbent named AbSolute. For this example, the optimum productivity was found to increase from 2.9 kg L⁻¹ day⁻¹ for batch operation to 4.0 kg L⁻¹ day⁻¹ for SMCC operation with three columns. A second case study considering a hypothetical adsorbent of larger particle size and slower mass transfer is also presented, to further demonstrate the applicability of the integrated approach. The case studies clearly illustrate the capabilities of the integrated approach in quickly determining the optimal design and operation for an SMCC arrangement and with minimal, carefully targeted, experimentation.     

Genetic Background of Fetal Growth Restriction

Fetal growth restriction (FGR) is one of the most formidable challenges in present-day antenatal care. Pathological fetal growth is a well-known factor of not only in utero demise in the third trimester, but also postnatal morbidity and unfavorable developmental outcomes, including long-term sequalae such as metabolic diseases, diabetic mellitus or hypertension. In this review, the authors present the current state of knowledge about the genetic disturbances responsible for FGR diagnosis, divided into fetal, placental and maternal causes (including preeclampsia), as well as their impact on prenatal diagnostics, with particular attention on chromosomal microarray (CMA) and noninvasive prenatal testing technique (NIPT).     

Effects of Enterococcus faecium (SF68) on immune function in mice

Probiotics exert health benefits on human and animals when administered in adequate amounts. The objective of this study was to assess the effects of Enterococcus faecium (SF68) on intestinal colonisation and immune function of BALB/c mice. Six-week-old female BALB/c mice were orally administered with E. faecium (SF68). Results showed that the total anaerobe and lactobacilli in the faeces increased (P < 0.05), while the number of faecal enterobacteria decreased (P < 0.05) in E. faecium-fed mice. Furthermore, supplementation of E. faecium (SF68) increased the percentage of double positive (DP) cells in peripheral blood, the concentration of plasma IgG, and the levels of interleukin-4 (IL-4), interleukin-6 (IL-6) and interferon-γ (IFN-γ) in splenocytes of the mice (P < 0.05). This study demonstrated that E. faecium SF68 affects the intestinal microbial flora and modulates the immune responses, which indicates a viable probiotic characteristic of E. faecium SF68 in modification of immune function.     

Fluctuation of Global Gene Expression by Endogenous miRNA Response to the Introduction of an Exogenous miRNA

Most of the intracellular endogenous microRNAs (endo-miRNAs) are considered to be saturated in Argonaute (Ago) proteins in the RNA-induced silencing complexes (RISCs). When exogenous miRNAs (exo-miRNAs) are introduced into cells, endo-miRNAs in the RISC may be replaced with exo-miRNAs or exo-miRNAs, and endo-miRNAs might also compete for the position in the newly synthesized RISC with each other. This would lead to the fluctuation of global gene expression not only by repression of exo-miRNA target gene expression, but also by the increase of the endo-miRNA target gene expression. In the present study, we quantified the changes in the expression levels of target genes of exo-miRNA and endo-miRNA in the cells transfected with fifteen different exo-miRNAs by microarray experiments. Different exo-miRNAs increased ratios of expression levels of target genes of a given endo-miRNA to different extents, suggesting that the replacement efficiencies might differ according to the exo-miRNA types. However, the increased ratios in the expression levels of each endo-miRNA target genes by the transfection of any particular exo-miRNA were mostly equivalent, suggesting that the endo-miRNAs present in the RISC might be replaced with excessive exo-miRNAs at similar levels, probably because they exist in single-stranded forms in the RISC.     

影响RNA芯片探针杂交效率的因素分析

RNA芯片作为研究ncRNA的主要技术之一,已受到广泛关注。然而因RNA芯片中靶序列的单链特征,使得RNA芯片的探针设计比DNA复杂。本研究对加强型绿色荧光蛋白（EGFP）RNA序列进行覆瓦式探针设计,制作了RNA原位合成芯片,研究探针／靶序列杂交双链△G和Tm、靶序列二级结构以及探针的末端碱基对RNA芯片杂交信号强度的影响。结果表明,探针／靶序列杂交双链△G和Tm相同的探针间的杂交信号存在较大差异,探针／靶序列杂交双链△G和,Tm与RNA芯片杂交信号之间未发现明显的规律性;靶序列二级结构的探针PT-sc参数与芯片杂交信号强度间呈较好的线性关系;探针5’末端碱基组成对芯片杂交信号强度也有影响,5’末端碱基为6／C的探针杂交信号总体上高于其邻近的5’末端碱基为A／T的探针的杂交信号,为RNA芯片的探针筛选提供一定的理论基础。     

蛋白质芯片微陈列条件优化及在疾病诊断上的应用

以PSA为模式抗体,对固定在氨基玻片表面时的甘油浓度、固定抗体的浓度、清洗缓冲溶液的选择等条件进行了优化。结果表明:含有30%(体积百分比)甘油的蛋白质溶液显示出灵敏度高、重现性好的微陈列点位;固定在氨基玻片表面的抗体的浓度较高时(2 mg.mL-1)可得荧光强度高的比较均匀的点位;依次用DI H2O、PBST、DI H2O洗涤氨基玻片表面时背景干扰少。按优化的蛋白质微陈列条件制备了芯片并尝试应用于对肿瘤标志物蛋白质PSA、AFP、CEA和CA15-3的检测。     

An introduction to DNA microarrays for gene expression analysis

This tutorial presents a basic introduction to DNA microarrays as employed for gene expression analysis, approaching the subject from a chemometrics perspective. The emphasis is on describing the nature of the measurement process, from the platforms used to a few of the standard higher-level data analysis tools employed. Topics include experimental design, detection, image processing, measurement errors, ratio calculation, background correction, normalization, and higher-level data processing. The objective is to present the chemometrician with as clear a picture as possible of an evolving technology so that the strengths and limitations of DNA microarrays are appreciated. Although the focus is primarily on spotted, two-color microarrays, a significant discussion of single-channel, lithographic arrays is also included.     

基于简单统计排名模型的差异表达基因识别

不同实验条件下差异表达基因(DEGs)的识别是微阵列数据分析的主要目标之一,针对分析结果中具有高排名的基因往往表现出较低差异表达水平的缺点,提出了一种基于简单统计排名模型的差异表达基因识别算法MRP(Matrix rank product)。算法可直接处理基因芯片原始数据,排除了数据预处理方法对算法的干扰;另外,通过对基因芯片数据形成的矩阵进行整体排序计算,得到具有高准确度的差异表达性排名结果。     

典型半潜式平台结构的微振子捕能特性研究北大核心CSCD

半潜式平台在海洋资源开发领域应用前景广阔,但存在平台设备能源供给不足的问题。本研究把常见的半潜式平台简化为三种典型的浮式基础,将波浪能发电装置集成至浮式基础,建立浮式基础与浮子耦合的运动方程,利用水动力仿真软件AQWA开展数值模拟和参数化分析,获得了波浪方向、浮式基础的截面形状、浮子直径以及浮子与浮式基础间距等因素对浮子捕能特性的影响规律。结果表明,四棱柱浮式基础在特定波浪方向下能够增强微阵列浮子的捕能效果,三棱柱浮式基础使微阵列浮子更好地适应各个方向的波浪激励,圆柱浮式基础以遮蔽效应为主,较大程度上减弱了浮子的捕能特性;不同波浪方向对四棱柱浮式基础、三棱柱浮式基础周围的浮子影响较大,而对基于圆柱浮式基础的微阵列浮子影响不大;通过增加浮子的直径以及浮子距圆柱浮式基础的距离,能够有效提高微阵列浮子平均输出功率。本研究可为半潜式平台与波浪能发电装置的集成提供参考。     

杜仲预防UVB致人成纤维细胞光老化的作用及基因水平的机制探讨

以UVB辐射体外培养的人成纤维细胞,探讨了杜仲乙醇提取物乙酸乙酯萃取部分(简称杜仲提取物)预防人成纤维细胞光老化的作用,并利用基因芯片对其可能的作用机制进行了探索。实验结果表明,杜仲提取物可以减少UVB对人成纤维细胞活性的影响,具有较好的预防光老化的作用,且基因芯片结果提示这种防护作用可能与提高细胞的抗氧化、DNA修复、抗凋亡有关,同时可能还具有一定促进细胞增殖和预防黑素细胞瘤的作用。