A schematic of the final analytical scheme is given in Fig. 1. All common chemicals were commercial analytical grade. Lysozyme selleck from chicken egg white, albumin from bovine serum (BSA), l-arabinose, glycogen from oyster, chondroitin sulfate

A sodium salt from bovine trachea, α-lactose monohydrate, glucose, N-acetyl neuraminic acid from Escherichia coli, Type II ι-carrageenan, 3-(N-morpholino)propanesulfonic acid (MOPS), and dextran were obtained from Sigma-Aldrich (United Kingdom). Deoxyribonucleic acid (DNA) sodium salt from salmon sperm was purchased from Fisher Bioreagents (United Kingdom). Sodium alginate from Laminaria hyperborea came from BDH (United Kingdom). Lonza Walkersville (Maryland, USA) provided endotoxin derived from E. coli. Gellan gum was produced by Applichem Biochemica (United Kingdom). High molecular weight hyaluronan (HA) was purchased from R&D Systems (United Kingdom). Endotoxin standards and stocks were purchased from Lonza Walkersville (Maryland, USA). All reagents were used as purchased. All sugars are D isomers except where noted otherwise. Solutions were filtered with 0.22 μm filters (GV PVDF, PES Express, Millipore, Massachusetts, USA) where appropriate. Polystyrene microtitre plates (Corning Costar #3596, Massachusetts, USA) were used with all assays, except where noted otherwise. All spectroscopic measurements were made with a Safire II spectrophotometer (Tecan,

Switzerland). Standard abbreviations this website for l-arabinose (Ara), ribose (Rib), glucose (Glc), galactose (Gal), glucuronic

acid (GlcA), guluronic acid (GulA), N-acetyl neuraminic acid (NeuNAc), mannose (Man) were selectively used to reduce graphical clutter. Absorbance spectra of pure carbohydrates and proteins were measured in solutions buffered with 20 mM MOPS, pH 7.2. Spectral measurements were also recorded following reaction in several colorimetric assays. Absorbance spectra were measured from 230 to 1000 nm in ≤3 nm increments using the microplate reader. The Bio-Rad protein assay (California, USA) based on Bradford’s method was employed to measure protein concentration [36] and [37]. The instructions provided by the reagent manufacturer (version: Lit 33 Rev C) were followed. Samples were diluted most in 20 mM MOPS, pH 7.0. Absorbance measurements were made at 595 nm and 990 nm. Blank-corrected standard curves were run in triplicate with absorbance at 990 nm subtracted from absorbance at 595 nm. Linear regression was used to fit the standard curve. The BCA assay kit (Pierce Thermo Scientific, Illinois, USA, version: 1296.7) was employed to measure protein concentration [40]. The microplate instructions provided by the assay kit manufacturer were followed. Samples were diluted into 20 mM MOPS, pH 7.0. Absorbance was measured at 562 nm and 990 nm. Blank-corrected standard curves were run in triplicate with a second order polynomial fit employed.