title: In situ generation of iminodiacetic acid groups on nanoporous alumina for the reversible immobilization of enzymes and other biomolecules.
abstract: Nanoporous alumina membranes were silanized with aminopropylsilane and iminodiacetic acid (IDA) groups were generated in situ by reaction with iodoacetate. The membranes were mounted in standard filter holders, connected to a HPLC system and saturated with selected metal ions. Cu(II) allowed the capture of chicken muscle lactate dehydrogenase with such stability, repeatability and reproducibility that Michaelis-Menten kinetics could be studied. The IDA surface was stable for months and could be depleted and regenerated with metal ions multiple times without appreciable loss of capacity. The binding of lactate dehydrogenase influenced the backpressure to the extent that could be expected for a monolayer according to Poiseuilles law.
authors.affiliation: Department of Chemistry-BMC, Uppsala University, Uppsala, Sweden.
authors.forename: Lennart
authors.initials: L
authors.lastname: Berglin
authors.forename: Marcus
authors.initials: M
authors.lastname: Kjellander
authors.forename: Gunnar
authors.initials: G
authors.lastname: Johansson
mesh_terms: D000537:Aluminum Oxide / Q000378:metabolism*; D000818:Animals; D002645:Chickens; D004795:Enzyme Stability; D004800:Enzymes, Immobilized / Q000378:metabolism*; D007098:Imino Acids / Q000378:metabolism*; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D009132:Muscles / Q000201:enzymology; D058608:Nanopores
chemical_list: D004800:Enzymes, Immobilized; D007098:Imino Acids; D007770:L-Lactate Dehydrogenase; D000537:Aluminum Oxide; C008109:iminodiacetic acid