Immunoaffinity chromatography of enzymes.
Immunoaffinity chromatography of enzymes represents an attractive purification technique suitable for one-step and large-scale purification of enzymes to homogeneity. Monoclonal and polyclonal antibodies can be used equally well. The broad use of the technique is restricted by the harsh elution conditions which are often required. The efforts to overcome these limitations and to optimize the method are reviewed, viz. proenzyme purification, purification of enzymes as part of multienzyme complexes carried out by a mild dissociation step, specific elution by substrates and effectors, enzyme stabilization, electrophoretical desorption and negative elution by adsorbing impurities from the crude extract, and hypotonic elution. The current practice is discussed considering antibody and enzyme selection, optimization of elution conditions, and washing steps using different media. Representative examples are given for various approaches.
Ehle
H
H
Institute of Biochemistry, Friedrich Schiller University Jena, GDR.
Horn
A
A
D000818:Animals; D002846:Chromatography, Affinity / Q000379:methods*; D004798:Enzymes / Q000302:isolation & purification*; D006801:Humans; D007163:Immunosorbent Techniques
D004798:Enzymes