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Recombinant aequorin with a reactive cysteine residue for conjugation with maleimide-activated antibody.

Analytical Biochemistry

immunology, chemistry, analysis, Recombinant Proteins, Molecular Sequence Data, Maleimides, Luminescent Measurements, genetics, Genetic Vectors, Cysteine, Biosensing Techniques, Antibodies, Amino Acid Sequence, Aequorin

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      Abstract

      The mutated recombinant aequorin with a reactive cysteine residue (Cys-aequorin) was highly purified and then conjugated with a maleimide-activated antibody without significant loss of luminescence activity. The conjugate ratio of Cys-aequorin to heavy chain of immunoglobulin G (IgG) was estimated to be 1:1. To test the bioluminescent immunoassay with aequorin-labeled antibody, alpha-fetoprotein (AFP), a serological marker of liver cancer, was used as a model analyte. The measurable range of AFP was 0.02 to 200 ng/ml with the coefficient of variation between 2.1 and 4.5%.

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      Journal
      10.1016/j.ab.2008.03.044
      18417074

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