FIELD: biochemistry.
SUBSTANCE: method for the isolation and/or purification of hemocyanin is described, which includes a) providing the composition of hemocyanin, obtained from marine mollusk, the specified composition is a serum hemolymph; b) reducing conductivity of the composition provided in a) stage to a value of 30 mS/cm to 10 mS/cm by adding dilution buffer; c) applying a diluted formulation containing hemocyanin obtained in b) stage onto an anion exchange chromatography column; d) rinsing said column with hemocyanin purification buffer, preferably to remove salts and other proteins; e) and eluting hemocyanin from said column by adding a second buffer. In addition, a method for the isolation and/or purification of an immunocyanin is presented, including: a) providing the composition of hemocyanin, obtained from marine mollusk, the specified composition is a serum hemolymph; b) reducing conductivity of the composition provided in a) stage to a value of 30 mS/cm to 10 mS/cm by adding dilution buffer; c) applying a diluted formulation containing hemocyanin obtained in b) stage onto an anion exchange chromatography column; d) rinsing said column with hemocyanin purification buffer, preferably to remove salts and other proteins; e) dissociating hemocyanin into subunits by adding dissociation buffer with pH from 8 to 10, not containing Ca++ and Mg++ ions; and f) eluting the immunocyanin from said column by adding a second buffer. Use of an anion exchange chromatography column is also disclosed containing a matrix material with a particle size of more than 30 mcm and a pore size of 3,000 to 8,000 A (Angstrem) for purifying hemocyanin or subunits thereof from hemolymph serum.
EFFECT: invention allows to isolate and/or purify hemocyanin and subunits thereof using anion-exchange chromatography.
12 cl, 6 ex
