1–3.5 pg/mL) and demonstrate an increased concentration of endogenous cytokines in disease, which were in keeping with mRNA expression data. These findings are consistent with published data on relative protein levels of these cytokines in Hp-infected and uninfected patients measured by ELISA, western blotting and Luminex in supernatants from gastric biopsy homogenate or gastric biopsy culture ( Bodger et al., 1997, Luzza et al., 2000, Shimizu et al., 2004, Mizuno et al., 2005 and Serelli-Lee et al., 2012). Sensitive measurement of cytokine selleck chemicals llc profiles using methodology that better reflects in vivo
concentrations is technically challenging. Optimisation of processing methods can improve data acquisition from precious tissue samples. A number of factors need to be considered when selecting an assay, including the type and quantity of samples, the availability and multiplexing capabilities of the desired analytes, the expected range of concentrations and sensitivity required, specificity, accuracy, precision, time and cost. We selected Luminex assays from MILLIPLEX for use in future studies based on our evaluation findings. Together with our optimised sample preparation protocol we concluded that Luminex assays are a suitable
technique for quantifying endogenous cytokines in mucosal biopsies. We hope that our approach will be more widely relevant for those seeking to quantify multiple cytokines in small tissue samples. The authors thank Dr Ian Spendlove, Dr Ann Lowe and Prof Jan Bradley for use of their Bio-Plex 200 systems, Dr Maria Toledo-Rodriguez for use of her CYTH4 TissueLyser LT, and the patients and staff at Nottingham University Vincristine chemical structure Hospital. We purchased all kits
used in this study. The study design, collection, analysis and interpretation of data, writing of the report and decision to submit for publication were undertaken independently by the authors without involvement of the funders or kit manufacturers. ES and RI are supported by Clinical Research Training Fellowships from the Medical Research Council [grant numbers G0701377 and G1000311]. This article presents independent research supported by the National Institute for Health Research (NIHR), through the NIHR Biomedical Research Unit in Gastrointestinal and Liver Diseases at Nottingham University Hospitals NHS Trust and the University of Nottingham. The views expressed are those of the author(s) and not necessarily those of the NHS, the NIHR or the Department of Health. “
“Several groups have attempted with varying degrees of success to improve bacterial production of antibody fragments by co-expressing them with molecular chaperones or folding catalysts (Bothmann and Pluckthun, 1998, Strachan et al., 1999, Bothmann and Pluckthun, 2000, Levy et al., 2001, Mavrangelos et al., 2001 and Maynard et al., 2005). The correct folding of scFv and Fab antibody fragments is highly dependent on the activity of peptidyl prolyl cis-trans isomerases (PPIases).