This project will investigate the use of Immobilized Metal Affinity Membranes for protein separation and recovery. A coupling chemistry will be developed for attaching IDA- iminodiacetate and TED-tris(carboxylmethyl) ethylenediamine with spacer arms to inorganic, porous glass microfiltration membranes. Loading isotherms for the ions Cu(II), Zn(II), and Ni(II) will be measured to determine binding constants and metal-ion stability during washing. The modified membranes will be used for fractionation of single and mixed proteins using model protein systems. Gradient and stepwise elution of adsorbed proteins will be attempted by varying pH, using displacers, and using a chelating agent. Immobilized metal affinity chromatography is developing into one of the most powerful techniques for purification of products from biotechnology. Rapid advances in membrane technology are also dramatically changing the engineering aspects of commercial scale biotechnology. The combination of these two techniques can have a potentially large effect on development of large-scale protein purification technologies.
