sepharose and Proteinuria

A case of a young adult with refractory nephrotic syndrome due to focal segmental glomerulosclerosis is reported. Several treatments had been used without success including steroids, cyclophosphamide, cyclosporine A, tacrolimus, and mycophenolate mofetil. Immunoadsorption was performed as a last resort to manage the nephrotic syndrome, which led to a drastic urinary protein reduction. We review the literature supporting immunoadsorption in primary focal segmental glomerulosclerosis.

Topics: Adult; Anemia, Megaloblastic; Angiotensin II Type 1 Receptor Blockers; Angiotensin-Converting Enzyme Inhibitors; Azathioprine; Blood Proteins; Combined Modality Therapy; Cyclophosphamide; Drug Resistance; Glomerulosclerosis, Focal Segmental; Humans; Immunosorbent Techniques; Immunosuppressive Agents; Male; Mycophenolic Acid; Nephrotic Syndrome; Plasmapheresis; Proteinuria; Sepharose; Staphylococcal Protein A; Tacrolimus

In prior work, a 50 kDa protein was purified to homogeneity from rat urine. This protein reduces food intake when injected into rats and is the only natural substance other than satietin known to be effective for long (24 hour) time periods and which does not make animals ill. However, when attempts were made to repeat the purification, contamination appeared in the 50 kDa fraction. The present contribution documents successful reisolation of the 50 kDa anorexigen by an improved method. Reisolation involved Cibacron blue-Sepharose, DEAE-Sephacel and Sephacryl S-200 chromatography, and SDS disc preparatory electrophoresis. The reisolated 50 kDa anorexigen contains no detectable carbohydrate. Partially purified preparations of the 50 kDa anorexigen were fragmented with trypsin and proteinase K without loss of anorexigenic activity. It is concluded that the 50 kDa anorexigen may be reproducibly purified to homogeneity and may contain within its amino acid sequence a peptide which is the basis of its anorexigenic activity.

Topics: Animals; Appetite Depressants; Biological Assay; Chemical Fractionation; Chromatography; Chromatography, Affinity; Chromatography, Gel; Coloring Agents; Eating; Electrophoresis, Polyacrylamide Gel; Endopeptidase K; Molecular Weight; Proteins; Proteinuria; Rats; Sepharose; Serine Endopeptidases; Trypsin

Many clinical laboratories do immunoelectrophoresis for qualitative assessment of proteins in biological fluids. Commercial kits are available that supply some or all of the necessary components, but the nature of these components varies. Seeking a reliable method for most easily preserving the original immunoelectrophoresis pattern as a permanent record, we compared a thick-film agar method to a thin-film agarose method in immunoelectrophoresis of a total of 70 serum and urine samples. For each sample, either method resulted in the same interpretation. The thin-film agarose method not only yields a stained, permanent record in about the same time that the thick film agar is ready for interpretation but not preservation, but also requires less antiserum, a smaller sample, and may not require purchase of additional hardware.

Topics: Blood Proteins; Humans; Immunoelectrophoresis; Proteinuria; Sepharose