Affinity Precipitation of (NAD Dependent Dehydrogenase) Lactate Dehydrogenase Using Cibacron Blue Dimers

Summary

The selective precipitation of lactate dehydrogenase, by using two structural variants of the Cibacron Blue dimers is demonstrated. While both the dimers are showing good selectivity, the dimer having a hexamethylene group as the bridge is more effective, both in terms of purification fold and yield. Thus, this dimer with hexamethylene bridge is able to precipitate selectively the lactate dehydrogenase from a crude extract of rabbit muscle with 110 fold purification, and 4750% yield, while a second dimer having a diaryl group gives a 90 fold purification with 2500% yield from the same extract. The purified enzyme shows a single band in the SDS-PAGE analysis.