Bioluminescent immunoassay with a protein A-luciferase fusion protein.
Protein A and firefly luciferase were genetically fused and the resulting fusion protein was applied to a bioluminescent immunoassay. The gene fusion plasmid, pMALU2, was constructed by inserting the structural gene of luciferase into a protein A expression vector, and was expressed in Escherichia coli. The resulting fusion protein of molecular weight 91 kDa retained not only the enzymatic activity of luciferase but also the binding capability of protein A to the Fc region of immunoglobulin G (IgG). The bioluminescent immunoassay was performed with the fusion protein and human IgG was determined in the concentration range from 10(-3) to 10(-7) g/ml.
Kobatake
E
E
Department of Bioengineering, Faculty of Bioscience and Biotechnology, Tokyo Institute of Technology, Yokohama, Japan.
Iwai
T
T
Ikariyama
Y
Y
Aizawa
M
M
D004926:Escherichia coli / Q000235:genetics; D005069:Evaluation Studies as Topic; D006801:Humans; D007124:Immunoenzyme Techniques; D007074:Immunoglobulin G / Q000032:analysis; D008156:Luciferases / Q000235:genetics; D008163:Luminescent Measurements; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000235:genetics; D013205:Staphylococcal Protein A / Q000235:genetics
D007074:Immunoglobulin G; D011993:Recombinant Fusion Proteins; D013205:Staphylococcal Protein A; D008156:Luciferases