Infrared absorbance spectroscopy of aqueous proteins : comparison of transmission and ATR data collection and analysis for secondary structure fitting

Attenuated total reflectance (ATR) infrared absorbance spectroscopy of proteins in aqueous solution is much easier to perform than transmission spectroscopy, where short path‐length cells need to be assembled reproducibly. However, the shape of the resulting ATR infrared spectrum varies with the refractive index of the sample and the instrument configuration. Refractive index in turn depends on the absorbance of the sample. In this work, it is shown that a room temperature triglycine sulfate detector and a ZnSe ATR unit can be used to collect reproducible spectra of proteins. A simple method for transforming the protein ATR spectrum into the shape of the transmission spectrum is also given, which proceeds by approximating a Kramers‐Krönig–determined refractive index of water as a sum of four linear components across the amide I and II regions. The light intensity at the crystal surface (with 45° incidence) and its rate of decay away from the surface is determined as a function of the wave number–dependent refractive index as well as the decay of the evanescent wave from the surface. The result is a single correction factor at each wave number. The spectra were normalized to a maximum of 1 between 1600 cm−1 and 1700 cm−1 and a self‐organizing map secondary structure fitting algorithm, SOMSpec, applied using the BioTools reference set. The resulting secondary structure estimates are encouraging for the future of ATR spectroscopy for biopharmaceutical characterization and quality control applications

Enhancing the recovery of oilseed rape seed oil bodies (oleosomes) using bicarbonate-based soaking and grinding media

An aqueous process for the recovery of oil bodies from rapeseed using sodium bicarbonate-based soaking and grinding media (pH 9.5) was investigated. The effect of the ratio between seed and mass of media during grinding and molarity of the medium used on oil body integrity, purity and storage stability have been studied. The grinding of seeds in solution at a ratio of 1:7 (w/w) significantly improved the quality of oil body suspension to a size more in-line with that seen in vivo (average D4,3 of 1.19 μm). The purity and the composition of the recovered oil bodies depends on the molarity of medium used; the use of a sodium bicarbonate solution (pH 9.5, 0.1 M) in the grinding and washing steps produced oil body preparations with the same purity as that resulting from washing a crude preparation with 9 M urea. The resultant emulsion had improved physical stability over a storage period of one month

Gas light and steam The gas works railways of the North Thames Gas Board and constituent companies

SIGLEAvailable from British Library Document Supply Centre- DSC:95/03789 / BLDSC - British Library Document Supply CentreGBUnited Kingdo