AI Article Synopsis
- The traditional method for isolating bovine dopamine-beta-hydroxylase (bDBH) resulted in a product that was active but contained about 50% impurities.
- To enhance purity, a specialized technique called immobilized metal chelate affinity chromatography was used, achieving over 90% pure and active bDBH.
- Additionally, the purification process included step elution with methyl-D-mannoside, and bDBH levels in different samples were analyzed using size-exclusion chromatography.
Article Abstract
The use of the traditional scheme for the isolation of bovine dopamine-beta-hydroxylase (bDBH) from bovine adrenal medulla resulted in active but not pure bDBH, containing about 50% of admixtures. Immobilized metal chelate affinity chromatography on agarose modified with iminodiacetic acid residues and charged with cobalt ions was applied in the final stage to obtain more than 90% pure and active bDBH. Final purification of bDBH using step elution with 0-0.5 M methyl-D-mannoside in buffer solution from concanavalin A-Sepharose was studied. The determination of bDBH in various samples was performed using size-exclusion chromatography.
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| http://dx.doi.org/10.1016/0021-9673(95)00179-q | DOI Listing |
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