Rapid screening of purification strategies for the capture of a human recombinant F(ab')2 expressed in baculovirus-infected cells using a micro-plate approach and SELDI-MS.

The development of a capture step of a human recombinant F(ab')(2) produced and expressed in baculovirus-infected cells was investigated by screening three mixed-mode chromatography sorbents (HEA HyperCel, PPA HyperCel and MEP HyperCel) and two ion exchangers (Q Ceramic HyperD F, S Ceramic HyperD F) sorbents using a 96-well plate format and SELDI-MS. HEA HyperCel gave the best separation performance therefore the conditions tested in micro-plate were transferred to laboratory scale chromatographic experiments, confirming that the recombinant F(ab')(2) was effectively captured on the mixed-mode sorbent without any pre-treatment of the crude extract with a 82% recovery and a 39-fold purification.

Effects of cadmium and uranium on some in vitro renal targets.

Metals are major pollutants not only in occupational settings but also in the general environment. Chronic exposure of workers has been related to severe damage, especially at the renal level. While toxic compounds such as metals are well known to severely impair tubular functions, it is clear that nephrotoxicants can act on various other renal targets, i.

Influence of cadmium speciation for the evaluation of in vitro cadmium toxicity on LLC-PK(1) cells.

The main objective of the present work was to assess the potentiality of in vitro models to improve our understanding of cadmium-induced toxicity, especially on epithelial renal cells. Indeed cadmium, a potent toxic metal, poses a serious environmental threat and the mechanisms of its renal toxicity need to be clarified. Cytotoxicity studies presented here were performed in a tubular proximal original established porcine kidney cell line (LLC-PK(1)).

Protective effects of polyphenols against cadmium-induced glomerular mesangial cell myocontracture.

The main objective of this work was to determine the ability of polyphenols (procyanidoloic oligomers; PCO) to diminish the contracture of CdCl2-induced mesangial cells (smooth muscle cell type). Glomeruli were isolated by passing rat renal cortex pulp through calibrated sieves followed by a culture step for outgrowth of cells. PCO lethality was measured by microassay (Neutral Red uptake).

Uranium-induced Vasoreactivity in Isolated Glomeruli and Cultured Rat Mesangial Cells.

The present study is aimed at investigating direct vasocontractive effects of uranium on two glomerular in vitro models (isolated rat glomeruli and cultured mesangial cells). For this study, sublethal doses of the uranium(VI)-bicarbonate complex were determined by cytotoxicity experiments on confluent cultured mesangial cells. Afterwards, using a morphoquantitative approach (computerized image analyser) the variations of planar surface areas (PSA) of glomeruli and mesangial cells exposed to uranium(VI)-bicarbonate have been assessed.