Visualisation of J-type counter-current chromatography: A route to understand hydrodynamic phase distribution and retention

This article can be accessed from the link below - Copyright @ 2012 Elsevier. The article was made available through open access by the Brunel Open Access Publishing Fund.This paper has addressed decade sought-after questions on phase bilateral distribution and stationary phase retention in any J-type high-speed counter-current chromatographic (CCC) centrifuge. Using a 2-D spiral column operated on such a CCC device and an aqueous two-phase system, this work systematically observed the phase interaction during transitional period and at dynamic equilibration under stroboscopic illumination. The experimental results thus obtained were used to examine the effects of the liquid–solid friction force, tangential centrifugal force, and physical properties of the two-phase system on hydrodynamic phase behaviour. We identified that (a) density difference between lower and upper phases is the critical factor to cause unusual phase bilateral distribution in the 2-D spiral column and (b) interfacial tension (manifested primarily as phase settling time) of any two-phase system is the critical factor in explaining inability to retain stationary phase in 3-D helical column and, for certain flow modes, in the 2-D spiral column. This work thus has extended or modified the well-established rule-of-thumb for operating J-type CCC devices and our conclusions can accommodate virtually all the anomalies concerning both hydrophobic and hydrophilic phase systems. To this end, this work has not only documented valuable experimental evidences for directly observing phase behaviour in a CCC column, but also finally resolved fundamentally vital issues on bilateral phase distribution orientation and stationary phase retention in 2-D spiral and 3-D helical CCC columns. Revised recommendations to end users of this technology could thus be derived out of the essence of the present work presumably following further experimental validation and a consensus in the CCC R&D and manufacturing circle.This work is funded by the BBSRC (UK) grant BB/C5063341/1, by MoST (China) as a Major State Basic Research Development Programme grant 2007CB714303 (973 Programme), by M & G Trading, and by Shanghai Science and Technology Board grant 11DZ2292700

Scale-up protein separation on stainless steel wide bore toroidal columns in the type-J counter-current chromatography

Manufacturing high-value added biotech biopharmaceutical products (e.g. therapeutic proteins) requires quick-to-develop, GMP-compliant, easy-to-scale and cost effective preparatory chromatography technologies. In this work, we describe the construction and testing of a set of 5-mm inner diameter stainless steel toroidal columns for use on commercially available preparatory scale synchronous J-type counter-current chromatography (CCC) machinery. We used a 20.2 m long column with an aqueous two-phase system containing 14% (w/w) PEG1000 and 14% (w/w) potassium phosphate at pH 7, and tested a sample loading of 5% column volume and a mobile phase flow rate of 20 ml/min. We then satisfactorily demonstrated the potential for a weekly protein separation and preparation throughput of ca. 11 g based on a normal weekly routine for separating a pair of model proteins by making five stacked injections on a single portion of stationary phase with no stripping. Compared to our previous 1.6 mm bore PTFE toroidal column, the present columns enlarged the nominal column processing throughput by nearly 10. For an ideal model protein injection modality, we observed a scaling up factor of at least 21. The 2 scales of protein separation and purification steps were realized on the same commercial CCC device.The BBSRC Grant number BB/FOF/206. YHG and YZ also acknowledge financial support by National Basic Research Program (973 Program 2013CB733600).Ms Yan Zhao (School of Mechanical and Power Engineering) has been grateful to a Training Program of Innovation and Entrepreneurship for Undergraduates Grant number X13219 by East China University of Science and Technology. Contribution and/or encouragement to this work from D. Fisher, T. Bunce, Ying-Dong Liu, Zheng-Tao Zhang, Li-Li Ba

Introduction to Special Issue - In-depth study of air pollution sources and processes within Beijing and its surrounding region (APHH-2 Beijing)

Abstract. The Atmospheric Pollution and Human Health in a Chinese Megacity (APHH-Beijing) programme is an international collaborative project focusing on understanding the sources, processes and health effects of air pollution in the Beijing megacity. APHH-Beijing brings together leading China and UK research groups, state-of-the-art infrastructure and air quality models to work on four research themes: (1) sources and emissions of air pollutants; (2) atmospheric processes affecting urban air pollution; (3) air pollution exposure and health impacts; and (4) interventions and solutions. Themes 1 and 2 are closely integrated and support Theme 3, while Themes 1-3 provide scientific data for Theme 4 to develop cost-effective air pollution mitigation solutions. This paper provides an introduction to (i) the rationale of the APHH-Beijing programme, and (ii) the measurement and modelling activities performed as part of it. In addition, this paper introduces the meteorology and air quality conditions during two joint intensive field campaigns - a core integration activity in APHH-Beijing. The coordinated campaigns provided observations of the atmospheric chemistry and physics at two sites: (i) the Institute of Atmospheric Physics in central Beijing, and (ii) Pinggu in rural Beijing during 10 November – 10 December 2016 (winter) and 21 May- 22 June 2017 (summer). The campaigns were complemented by numerical modelling and automatic air quality and low-cost sensor observations in the Beijing megacity. In summary, the paper provides background information on the APHH-Beijing programme, and sets the scene for more focussed papers addressing specific aspects, processes and effects of air pollution in Beijing

Finishing the euchromatic sequence of the human genome

The sequence of the human genome encodes the genetic instructions for human physiology, as well as rich information about human evolution. In 2001, the International Human Genome Sequencing Consortium reported a draft sequence of the euchromatic portion of the human genome. Since then, the international collaboration has worked to convert this draft into a genome sequence with high accuracy and nearly complete coverage. Here, we report the result of this finishing process. The current genome sequence (Build 35) contains 2.85 billion nucleotides interrupted by only 341 gaps. It covers ∼99% of the euchromatic genome and is accurate to an error rate of ∼1 event per 100,000 bases. Many of the remaining euchromatic gaps are associated with segmental duplications and will require focused work with new methods. The near-complete sequence, the first for a vertebrate, greatly improves the precision of biological analyses of the human genome including studies of gene number, birth and death. Notably, the human enome seems to encode only 20,000-25,000 protein-coding genes. The genome sequence reported here should serve as a firm foundation for biomedical research in the decades ahead