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PubMed · 7841768

Fluidized-bed receptor-affinity chromatography.

C Spence·C A Schaffer·S Kessler·P Bailon
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Abstract

A multipurpose fluidized-bed receptor-affinity purification system based upon the biological recognition between an immobilized receptor and its soluble protein ligands is described. The fluidized affinity sorbent consists of a soluble form of interleukin-2 receptor chemically bonded to an aldehyde derivative of controlled pore glass beads, which have a pore diameter of 1000 A and a particle density of 1.2-1.3 g/mL. The fluidized-bed separation device used in this study consists of a specially designed column fitted at the inlet end with a perforated distributor plate covered with a screen and the top outlet with an adjustable piston. The fluidized-bed consisting of a loose gel matrix permits the unimpeded passage of cell debris and particulate matter, while the target protein is captured by the affinity beads. Purification of the humanized-anti-Tac monoclonal antibody is used as a model system to determine the operational parameters. Also, fluidized-bed receptor-affinity chromatography has been successfully employed in the purification of recombinant interleukin-2 and single chain anti-Tac(Fv)-Pseudomonas exotoxin immunotoxin from unclarified inclusion body extracts. Overall, fluidized-bed receptor-affinity chromatography is found to be a productive affinity method suitable for the purification of recombinant human interleukin-2 and related molecules.

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ADP Ribose TransferasesAntibodies, MonoclonalBacterial ToxinsCell LineChromatography, AffinityExotoxinsHumansImmunotoxinsInterleukin-2Receptors, Interleukin-2Recombinant ProteinsVirulence FactorsPseudomonas aeruginosa Exotoxin AJournal Article

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C Spence, C A Schaffer, S Kessler, P Bailon. Fluidized-bed receptor-affinity chromatography.. https://doi.org/10.1002/bmc.1130080508

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