Mammalian cell production of a respiratory syncytial virus (RSV) candidate vaccine recovered using a product-specific affinity column

The recombinant production of a respiratory syncytial virus (RSV) candidate vaccine BBG2Na in baby hamster kidney cells (BHK-21 cells) was investigated. BBG2Na consists of a serum-albumin-binding region (BB) fused to a 101-amino-acid fragment of the RSV G-protein. Semliki Forest virus-based expression vectors encoding both intracellular and secreted forms of BBG2Na were constructed and found to be functional. Affinity recovery of BBG2Na employing human serum albumin columns was found to be inefficient due to the abundance of BSA in the applied samples. Instead, a strategy using a tailor-made affinity ligand based on a combinatorially engineered Staphylococcus aureus protein A domain, showing specific binding to the G-protein part of the product, was evaluated. In conclusion, a strategy for production and successful recovery of BBG2Na in mammalian cells was created, through the development of a product-specific affinity column.

Hydroisomerisation of n-alkanes over partially reduced MoO3: Promotion by CoAlPO-11 and relations to reaction mechanism and rate-determining step

The reaction mechanism and the rate-determining step (RDS) of the isomerisation of n-alkanes (C-4-C-6) over partially reduced MoO3 catalysts were studied through the effects of the addition of an alkene isomerisation catalyst (i.e. CoAlPO- 11). When an acidic CoAlPO- 11 sample was mechanically mixed with the MoO3, a decrease of the induction period and an increase of the steady-state conversion of n-butane to isobutane were observed. These data support previous assumptions that a bifunctional mechanism occurred over the partially reduced MoO3 (a complex nanoscale mixture of oxide-based phases) during n-butane isomerisation and that the RDS was the skeletal isomerisation of the linear butene intermediates. The only promotional effect of CoAlPO-11 on the activity of partially reduced MoO3 for C-5-C-6 alkane hydroisomerisation was a reduction of the induction period, as the RDS at steady-state conditions appeared to be dehydrogenation of the alkane in this case. However, lower yields of branched isomers were observed in this case, the reason of which is yet unclear. (c) 2005 Elsevier B.V. All rights reserved.

Purification of imidazolium ionic liquids for spectroscopic applications

Ionic liquids are often contaminated by colored impurities. These impurities can be problematic for spectroscopic studies or for monitoring organic reactions by UV/Vis spectroscopy. The effect of different purification methods on the optical quality of colored ionic liquids was studied and compared. Yellowish ionic liquids can partially be decolorized by treatment with active charcoal or by recrystallization. Our experiments show column liquid chromatography is not always a good technique to prepare spectrograde imidazolium halide ionic liquids. Colorless and UV-transparent ionic liquids were synthesized by a method that can exclude the need for further purification steps. (c) 2005 Elsevier B.V. All rights reserved.

The settlement performance of stone column foundations

Vibrated stone columns are frequently used as a method of reinforcing soft ground as they provide increased bearing capacity and reduce foundation settlements. Their performance in relation to bearing capacity is well documented, but there is also a need for enhanced understanding of their settlement characteristics, particularly in relation to small-group configurations. This paper presents results obtained from physical model tests on triaxial specimens 300 mm in diameter and 400 mm high. Parameters investigated include column length to diameter ratio, area replacement ratio and single/group configuration. The findings of the work are as follows. The design is flexible: settlement can equally be controlled using short columns at relatively high area replacement ratios, or longer columns at smaller area replacement ratios. An optimum area replacement ratio of 30-40% exists for the control of settlement. The settlement performance of a small column group is highly influenced by inter-column and footing interaction effects.

The pressure distribution along the stone column in soft clay when subjected to consolidation and foundation loading

The mechanism whereby the foundation loading is transmitted through stone the column (included in soft clay) has received less attention from researchers. This paper reports on some interesting findings obtained from a laboratory-based model study in respect of this issue. The stone column, included in the soft clay bed was subjected to foundation loading under drained conditions. The results show, probably for the first time, how the foundation loadings are transmitted through the column and indeed the existence of “negative skin friction” (a widely accepted phenomena in solid piles) in granular columns in soft clays.

Detection of epidermal growth factor receptor variations by partially denaturing HPLC

Background: Epidermal growth factor receptor gene (EGFR) variants may be useful markers for identifying responders to gefitinib and erlotinib, small-molecule tyrosine kinase inhibitors of EGFR; therefore, sensitive and cost-effective assays are needed to detect EGFR variants in routine clinical samples. We have developed a partially denaturing HPLC (pDHPLC) assay that is superior to direct sequencing with respect to detection limits, costs, and time requirements.

Effect of column base strength on steel portal frames in fire

This paper presents the findings of a project part sponsored by an ICE Research and Development grant on portal frames in fire. The research reported here has also lead to a sucessful research grant from the IStructE. The paper describes a non-linear elasto plastic dynamic finite element model that captures the collapse of a portal frame in fire. It demonstrates that current guidance on the column base stiffness and strength, to prevent collapse, may in some cases be unconservative.