Protein adsorption on histidyl-aminohexyl-Sepharose 4B. II. Application to the negative one-step affinity purification of human beta2-microglobulin and immunoglobulin G.
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.
Pitiot
O
O
Laboratoire d'Interactions Moléculaires et de Technologie des Séparations, Université de Technologie de Compiègne, CNRS-UPRESA 6 022, France.
Nedonchelle
E
E
Legallais
C
C
Vijayalakshmi
M A
MA
D000327:Adsorption; D002846:Chromatography, Affinity / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007074:Immunoglobulin G / Q000097:blood / Q000302:isolation & purification*; D011506:Proteins / Q000737:chemistry*; D012685:Sepharose / Q000737:chemistry*; D021241:Spectrometry, Mass, Electrospray Ionization; D014511:Uremia / Q000097:blood; D001613:beta 2-Microglobulin / Q000097:blood / Q000302:isolation & purification*
D007074:Immunoglobulin G; D011506:Proteins; D001613:beta 2-Microglobulin; D012685:Sepharose