The general potential of biospecific affinity chromatography in enzyme isolation and purification is well recognised. In the present study, three distinct approaches were assessed for the rapid, one-chromatographic-step affinity purification of L-lactate dehydrogenase (L-LDH, EC 1.1.1.27) from bovine heart: (i) the specific ligand approach using an immobilised oxamate derivative; (ii) the general ligand approach (immobilised NAD(+) derivatives) in conjunction with a kinetic 'locking-on' strategy; (iii) the general ligand approach in conjunction with the kinetic locking-on strategy and an auxiliary tactic (the 'stripping' ligand tactic). Purification tables were constructed for each of the purification protocols and all purified enzyme samples were assessed for homogeneity using SDS-PAGE. Results confirm that the more widely applicable general ligand approach, when used in conjunction with both the locking-on strategy and stripping ligand tactic, has the potential to purify NAD(+)-dependent dehydrogenases to homogeneity in a single bioaffinity chromatographic step.