sepharose and Uremia

The adsorption of two human proteins, beta2-microglobulin and Immunoglobulin G, from uremic patient's blood ultrafiltrate and plasma, respectively, was investigated on the histidyl-aminohexyl-Sepharose 4B adsorbent. Both target proteins could be adsorbed on the gel through a low affinity for immobilized histidine ligand. However, a fine adjustment of the operating conditions (ionic strength, buffer, pH) prevented their adsorption and thus allowed their "negative affinity" purification (purity estimated by silver nitrate SDS-PAGE) by the removal of the contaminating proteins. This simple and efficient method provides purification under gentle chromatographic conditions and a further characterization of both molecules.

Topics: Adsorption; beta 2-Microglobulin; Chromatography, Affinity; Electrophoresis, Polyacrylamide Gel; Humans; Hydrogen-Ion Concentration; Immunoglobulin G; Proteins; Sepharose; Spectrometry, Mass, Electrospray Ionization; Uremia

We evaluated the effect of azotemia on results for glycated hemoglobin as measured by a boronate-agarose affinity method and an ion-exchange chromatographic procedure with saline preincubation and found a good correlation. However, values for glycated hemoglobin in samples from nondiabetic patients with various degrees of azotemia were consistently higher with the ion-exchange column procedure (mean, 8.5%) than with the boronate affinity method (mean, 6.2%). The latter method may thus be preferred for monitoring glycated hemoglobin in diabetic patients with impaired renal function.

Topics: Boronic Acids; Chromatography, Affinity; Chromatography, Ion Exchange; Diabetes Mellitus; False Positive Reactions; Gels; Glycated Hemoglobin; Humans; Sepharose; Uremia

Topics: Humans; Immunity, Cellular; In Vitro Techniques; Leukocyte Migration-Inhibitory Factors; Leukocytes; Lymphokines; Sepharose; Uremia