Use of proZZ-obelin fusion protein in bioluminescent immunoassay.
Obelin is a photoprotein that emits light by Ca2+-binding. To develop a bioluminescent immunoassay based on the light emission property of obelin, we have expressed the apoobelin fusion protein with ZZ-domain of S. aureus protein A in E. coli by recombinant DNA techniques. The proZZ-obelin expressed was purified by one-step affinity chromatography on IgG-Agarose. The purified proZZ-obelin has both the luminescent activity of obelin and the IgG-binding ability of ZZ-domain. The specific activity of fusion protein was 8.5 X 10(15) photons per mg of protein.
Frank
L A
LA
Institute of Biophysics, Russian Academy of Sciences, Siberian Branch, Krasnoyarsk, Russia.
Illarionova
V A
VA
Vysotski
E S
ES
D000818:Animals; D000907:Antibodies, Bacterial / Q000032:analysis*; D002118:Calcium / Q000378:metabolism*; D004926:Escherichia coli; D006801:Humans; D007074:Immunoglobulin G / Q000032:analysis*; D007136:Immunoglobulins / Q000032:analysis*; D007700:Kinetics; D008163:Luminescent Measurements; D008164:Luminescent Proteins / Q000302:isolation & purification / Q000378:metabolism; D051379:Mice; D010957:Plasmids; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000302:isolation & purification / Q000378:metabolism; D015183:Restriction Mapping; D014376:Tuberculosis / Q000276:immunology*
D000907:Antibodies, Bacterial; D007074:Immunoglobulin G; D007136:Immunoglobulins; D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; C010497:obelin; D002118:Calcium