Evaluation of Immobilized Metal-Ion Affinity Chromatography (IMAC) as a Technique for IgG1 Monoclonal Antibodies Purification: The Effect of Chelating Ligand and Support

Monoclonal antibodies (MAbs) have been used for therapies and some analytical procedures as highly purified molecules. Many techniques have been applied and studied, focusing on monoclonal antibodies purification. In this study, an immobilized metal affinity chromatography membrane was developed and evaluated for the purification of anti-TNP IgG₁ mouse MAbs from cell culture supernatant after precipitation with a 50% saturated ammonium sulfate solution. The chelating ligands iminodiacetic acid, carboxymethylated aspartic acid (CM-Asp), nitrilotriacetic acid, and tris (carboxymethyl) ethylenediamine in agarose gels with immobilized Ni(II) and Zn(II) ions were compared for the adsorption and desorption of MAbs. The most promising chelating ligand—CM-Asp—was then coupled to poly(ethylene vinyl alcohol) (PEVA) hollow fiber membranes. According to SDS-PAGE and ELISA analyses, a higher selectivity and a purification factor of 85.9 (fraction eluted at 500 mM Tris) were obtained for IgG₁ using PEVA-CM-Asp-Zn(II). The anti-TNP MAb could be eluted under mild pH conditions causing no loss of antigen binding capacity.     

Molecular control of recombination dynamics in dye sensitised nanocrystalline TiO2 films

Modification of the structure of a porphyrin dye shows a significant change in the rate of charge recombination between injected electrons in the TiO2 and the oxidized dye anchored to it following optical excitation, offering an insight into fundamental understanding of processes occurring at the dye/semiconductor interface.     

Double-pass imaging polarimetry in the human eye

A Mueller-matrix imaging polarimeter was developed to measure spatially resolved polarization properties in the living human eye. The apparatus is a double-pass setup that incorporates two liquid-crystal variable retarders and a slow-scan CCD camera in the recording stage. Series of 16 images for the combinations of independent polarization states in the first and second passages were recorded for two experimental conditions: with the camera conjugated either with the retina or with the eye's pupil plane. Spatially resolved collections of Mueller matrices and the degree of polarization were calculated from those images for both retinal and pupil planes.     

Complexes containing phosphorus lignads—161: New phosphinite,phosphonite and phosphite derivatives of ruthenium,osmium and iridium

The complexes [MHCl(CO)(PPh₃)₃] (M = Ru or Os) readily undergo substitution at the site trans to the hydride ligand to afford phosphinite-, phosphonite-, or phosphite-containing products [MHCI(CO)(PPh₃)₂L] [L = P(OR)Ph₂, P(OR)₂Ph or P(OR)₃ respectively; R = Me or Et]. The ruthenium complexes alone undergo further substitution to afford complex cations [RuH(CO)(PPh₃)_nL_4?n]⁺ [n = 2, L = P(OMe)₃; n = 1, L = P(OR)₃; n = 0, L = P(OR)₂Ph or P(OR)Ph₂] which were isolated and characterised as their tetraphenylborate salts. Synthesis of the cationic complexes [IrHL₅][BPh₄]₂ [L = P(OR)₃, R = Me or Et] is also reported. Stereochemical assignments based on NMR data are given, and second order ³¹P and high field ¹H NMR patterns are analysed.