Integrated removal of nucleic acids and recovery of LDH from homogenate of beef heart by affinity precipitation

Authors: Dissing, Ulla; Mattiasson, Bo

Source: Bioseparation, Volume 7, Numbers 4-5, July 1998 , pp. 221-229(9)

Publisher: Springer

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Abstract:

Lactate dehydrogenase (LDH) was purified from beef heart homogenate by affinity precipitation. The protein purification was integrated with nucleic acid removal and was done by precipitation of nucleic acids by addition of poly(ethylene imine) PEI onto which a ligand, Cibacron blue, had been coupled. The yield of LDH after elution from the precipitate was 63%, the purification factor 6.9 and the nucleic acid content was reduced by 98%. The capacity of the affinity polymer Cibacron blue-PEI is dependent on the nucleic acid concentration in the homogenate. The beef heart homogenate had an unfavourable ratio of nucleic acids to LDH. Precipitation with recirculated Cibacron blue-PEI, already complexed with some nucleic acids, improved the yield of the enzyme to 74%. The loss of Cibacron blue-PEI, when recirculated, was less than 1% after each cycle.

Keywords: affinity precipitation; LDH; nucleic acids; poly(ethylene imine)

Document Type: Research article

DOI: http://dx.doi.org/10.1023/A:1008003701018

Publication date: 1998-07-01