Practical application of aqueous two-phase partition to process development for the recovery of biological products

The practical application of aqueous two-phase systems (ATPS) to process development has been exploited for several years for the recovery of biological products. Unfortunately, this has not resulted in an extensive presence of the technique in commercial processes. Some of the main identified reasons for such situation involve the full understanding of the mechanism governing phase formation and the behaviour of solute partitioning in ATPS processes, the cost of phase forming polymers and the necessary extended time to understand the technique for process development. In this review paper, some of the practical disadvantages attributed to ATPS are addressed. The practical approach exploited to design ATPS processes, the application to achieve process integration, the increasing use for the recovery of high-value products and the recent development of alternative low cost ATPS, are discussed. It is proposed that the potential trend in the application of ATPS processes for the recovery of biological products will involve the recovery of high-value bio-particulate products with medical applications. This proposed trend in the application of ATPS will address the urgent need to rapidly and economically bring new biopharmaceutical products to market using scaleable and efficient bioprocess technology.     

流程管理对高龄全髋关节置换术患者的临床效果分析

目的：随着我国人口老龄化日渐严重,高龄人群中髋关节骨折或坏死的发病率明显呈上升趋势。目前,临床用于治疗该类疾病的方法主要是全髋关节置换术（THA）。但高龄患者身体机能出现衰退,术后极易出现并发症。因此,高龄患者行全髋关节置换术的围术期护理工作起着重要的作用。本文针对行全髋关节置换术的高龄患者,探讨流程管理模式的临床效果,为护理工作提供参考。方法：选择我科2010年12月至2013年6月行全髋关节置换术的12位高龄患者,随机分为对照组和观察组,分别采用常规模式和流程管理模式进行护理。对比两组患者的临床效果及对护理服务的满意度。结果：观察组患者无一例并发症出现,髋关节功能恢复良好,均痊愈出院,且住院时间比对照组短（P〈0．05）;观察组对护理服务的满意度高于对照组,差异具有统计学意义（P〈0．05）。结论：采取流程管理模式对高龄患者的围术期护理具有显著的效果,有助于减少术后并发症,提高患者对护理服务的满意度,值得临床推广。     

Single-chain antibody fragment production in Pichia pastoris: Benefits of prolonged pre-induction glycerol feeding

Secretory production of a single-chain antibody fragment (scFv) by recombinant Pichia pastoris using the methanol inducible AOX1 promoter is limited biochemically by retarded secretion, and economically by the high demand for pure oxygen. To address the problem, the adaptation phase with growth-limiting feeding of glycerol before the production phase was optimized. In a standard procedure with a short glycerol-feeding phase before induction, scFv accumulated in the supernatant only after 15 h. Conversely, scFv started to appear immediately in the medium upon methanol induction when the glycerol-feeding phase was extended to 18 h. Interestingly, despite a significantly lower cell density in the cultivation with extended glycerol feeding, the same amount of functional product of 300 mg/L was obtained about 30 h after the start of glycerol feeding with both methods. mRNA analysis revealed that the higher and faster production of the product was related to longer lasting induction of the scFv mRNA. Additional effects of a better adaptation of the secretion machinery may be suggested by higher expression of unfolded protein response-related genes KAR2 and PDI. A clear benefit of the longer glycerol-feeding phase was a 75% reduction of the consumption of both pure oxygen and methanol, and a significantly lower cell density, which would be beneficial for down-stream purification of the product.     

预添加乳酸对直投式发酵泡萝卜感官品质的影响

以红萝卜为原料,在新配制发酵液时预先添加少量不同浓度梯度的乳酸,通过分析泡萝卜感官评价、各品质指标的动态变化规律以及有机酸和游离氨基酸测定结果,研究在直投式发酵下,经乳酸添加后的泡萝卜发酵过程以及风味品质差异,从而改进我国规模化的工业化泡菜生产工艺。结果表明,乳酸的添加能有效加快泡萝卜颜色、质地变化,缩短盐度、可溶性总糖、总酸含量达到平衡的时间,有机酸、游离氨基酸种类、含量均优于对照组泡萝卜,处理组泡萝卜整体成熟度更高。并最终结合感官评分结果确定出0.45%为最优乳酸添加量,所发酵出的泡萝卜感官品质最佳。     

Process control in cell culture technology using dielectric spectroscopy

In the biopharmaceutical industry, mammalian and insect cells as well as plant cell cultures are gaining worldwide importance to produce biopharmaceuticals and as products themselves, for example in stem cell therapy. These highly sophisticated cell-based production processes need to be monitored and controlled to guarantee product quality and to satisfy GMP requirements. With the process analytical technology (PAT) initiative, requirements regarding process monitoring and control have changed and real-time in-line monitoring tools are now recommended. Dielectric spectroscopy (DS) can serve as a tool to satisfy some PAT requirements. DS has been used in the medical field for quite some time and it may allow real-time process monitoring of biological cell culture parameters. DS has the potential to enable process optimization, automation, cost reduction, and a more consistent product quality. Dielectric spectroscopy is reviewed here as a tool to monitor biochemical processes. Commercially available dielectric sensing systems are discussed. The potential of this technology is demonstrated through examples of current and potential future applications in research and industry for mammalian and insect cell culture.     

Sludge exchange process on two serial CSTRs anaerobic digestions: process failure and recovery

The sludge exchange process using two anaerobic digesters (CSTRs) in series was investigated under the mesophilic condition (36-38 °C). At first, the digesting sludge of the CSTRs in series with different TVFA/alkalinity ratios was tested in the laboratory by mixing the digesting sludge of two CSTRs from 6.5% to 50% based on volume. The sludge exchange test was then performed using the same CSTRs under batch and continuous processes. The change in the TVFA/alkalinity ratio was found to be linear with the digesting sludge exchange volume. The CSTR of TVFA/alkalinity ratio 1.970 recovered completely failed within 11 days for the batch process and the CSTR of TVFA/alkalinity ratio 1.514 within 3 weeks for the continuous feeding process at a sludge exchange volume of 13%. The reactor operation was stable when the TVFA/alkalinity ratio was less than 1.0 and when the TVFA concentration was lower than 10,000 mg L⁻¹.     

On-field study of anaerobic digestion full-scale plants (Part II): new approaches in monitoring and evaluating process efficiency

Biogas plants need easy and practical tools for monitoring and evaluating their biological process efficiency. As soon as, in many cases, biomass supply present considerable costs, full-scale anaerobic digestion (AD) processes must approach, as much as possible, the potential biogas yield of the organic mixture fed to the biodigesters. In this paper, a new indicator is proposed (the bio-methane yield, BMY), for measuring the efficiency in full-scale AD processes, based on a balance between the biochemical methane potential (BMP) of the input biomass and the residual BMP of the output materials (digestate). For this purpose, a one-year survey was performed on three different full-scale biogas plants, in the Italian agro-industrial context, and the bio-chemical processes were fully described in order to calculate their efficiencies (BMY = 87-93%) and to validate the new indicator proposed, as useful and easily applicable tool for full-scale AD plants operators.     

Characterization of photosynthetically active duckweed (<Emphasis Type="Italic">Wolffia australiana</Emphasis>) in vitro culture by Respiration Activity Monitoring System (RAMOS)

The feasibility of oxygen transfer rate (OTR) measurement to non-destructively monitor plant propagation and vitality of photosynthetically active plant in vitro culture of duckweed (Wolffia australiana, Lemnaceae) was tested using Respiration Activity Monitoring System (RAMOS). As a result, OTR proofed to be a sensitive indicator for plant vitality. The culture characterization under day/night light conditions, however, revealed a complex interaction between oxygen production and consumption, rendering OTR measurement an unsuitable tool to track plant propagation. However, RAMOS was found to be a useful tool in preliminary studies for process development of photosynthetically active plant in vitro cultures.     

Sustainability in the biopharmaceutical industry: Seeking a holistic perspective

Biopharmaceuticals manufacturing is a critical component of the modern healthcare system, with emerging new treatments composed of increasingly complex biomolecules offering solutions to chronic and debilitating disorders. While this sector continues to grow, it strongly exhibits “boom-to-bust” performance which threatens its long-term viability. Future trends within the industry indicate a shift towards continuous production systems using single use technologies that raises sustainability issues, yet research in this area is sparse and lacks consideration of the complex interactions between environmental, social and economic concerns. The authors outline a sustainability-focused vision and propose opportunities for research to aid the development of a more integrated approach that would enhance the sustainability of the industry.