ISSN 1995-5537

“Biotechnology” journal Т. 1, № 1, 2008
P. 100-105, Bibliography. 5, Ukraininan.
Universal Decimal classification:  616.36:578.891

Highly effective technology for production of recombinant analogue of p24 protein of bovine leukemia virus with proven antigenic properties using immunoenzyme was developed. During investigation the best strain E. coli BL21(DE3) CodonPlus-RIL for introduction of recombinant plasmid was chosen, the composition of growth media was optimized. The procedure of metal-helate-affinity chromatography and refolding of recombinant protein was developed. The results of immunoenzyme were used as criteria of quality of recombinant protein on each stage of the work. The influence of bacterial host strains and growth media composition on the yield and distribution of recombinant protein between soluble form in cytoplasm and insoluble inclusion bodies was shown. Using this technology it is possible to obtain recombinant protein with 98% purity and improved antigenic properties. This protein can be used to increase specificity and sensitivity of BLV-diagnostic test kits based on results of immunoenzyme.

Key words: bovine leukemia virus, protein p24, Escherichia coli BL21 (DE3), metal-helate affinity chromatography, refolding, enzyme-linked immunosorbent assay