Enzyme Leakage and Multipoint Attachment of Agarose‐Bound Enzyme Preparations
Open Access
- 31 December 1977
- journal article
- research article
- Published by Wiley in European Journal of Biochemistry
- Vol. 82 (1), 181-186
- https://doi.org/10.1111/j.1432-1033.1978.tb12010.x
Abstract
The solvolytic detachment of leucine aminopeptidase fromSepharose-enzyme conjugates with multiple and single anchoring bonds has been studied under a variety of conditions by radiochemical and enzymological methods. The release of the single-point-fixed conjugate could be described by a leakage function, derived previously, yielding the first-order rate constant of the cleavage of the enzyme-matrix bond. The nucleophile hydroxylamine increased the detachment rate considerably. The release of the immobilized enzyme was incomplete in all experiments even after prolonged times. The enzyme leakage from multipoint-attached conjugates was still high enough to prohibit a long-term use of such preparations in routine work at room temperature.This publication has 19 references indexed in Scilit:
- Studies on conformation of soluble and immobilized enzymes using differential scanning calorimetry. 2. Specific activity and thermal stability of enzymes bound weakly and strongly to Sepharose CL 4BBiochemistry, 1977
- Immobilization of enzymes by covalent binding to amine supports via cyanogen bromide activationBiochemical and Biophysical Research Communications, 1976
- Application of spectrophotometric methods to the determination of protein bound to agarose beadsAnalytical Biochemistry, 1975
- Ligand-leakage in affinity chromatography, a mathematical approachCellular and Molecular Life Sciences, 1974
- Stable, high capacity and non charged agarose derivatives for immobilization of biologically active compounds and for affinity chromatographyMolecular and Cellular Biochemistry, 1974
- Limitations of Affinity Chromatography: Solvolytic Detachment of Ligands from Polymeric SupportsHelvetica Chimica Acta, 1974
- Stable and high capacity Sepharose derivatives for affinity chromatographyFEBS Letters, 1973
- Limitations of affinity chromatography: Sepharose‐bound cyclic 3′,5′‐adenosine monophosphateFEBS Letters, 1972
- Chemical Coupling of Enzymes to Cross-Linked Dextran (‘Sephadex’)Nature, 1966
- Die Kupplung von Proteinen an diazotiertes PolyaminostyrolHoppe-Seyler´s Zeitschrift Für Physiologische Chemie, 1954