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A Recombinant scFv/Streptavidin-Binding Peptide Fusion Protein for the Quantitative Determination of the Scorpion Venom Neurotoxin AahI by Nicolas Aubrey; Christiane Devaux; Eric di Luccio; Max Goyffon; Hervé Rochat; Philippe Billiald is a Biochemistry, Genetics and Molecular Biology article available to read on EtoBox.

What is A Recombinant scFv/Streptavidin-Binding Peptide Fusion Protein for the Quantitative Determination of the Scorpion Venom Neurotoxin AahI about?

We created a construct encoding a peptide known to mimic the binding properties of biotin fused to the carboxy-terminus of a scFv fragment that binds a scorpion toxin (AahI). This fusion protein was produced in the periplasm of bacteria and purified to homogeneity by single-step affinity chromatography on streptavidin-agarose with a yield close to 1 mg/l. DNA sequencing, dot blot and mass spectrometric analyses demonstrated the integrity of the soluble immunoconjugate. Fusion to the streptavidin-binding peptide did not affect the ability of the scFv to recognize its antigen with a high affinity (K d = 2.3 × 10 -10 M). Similarly, the streptavidin-binding property was not impaired in the fusion protein. Thus, the immunoconjugate was bifunctional and had a low molecular mass of 28 kDa. This enabled us to develop rapid and sensitive immunoassays for the specific detection of the toxin AahI accurately to 0.6 ng/ml, opening up new perspectives for the diagnosis of envenomations.

Who reads A Recombinant scFv/Streptavidin-Binding Peptide Fusion Protein for the Quantitative Determination of the Scorpion Venom Neurotoxin AahI?

Author

Nicolas Aubrey; Christiane Devaux; Eric di Luccio; Max Goyffon; Hervé Rochat; Philippe Billiald

Publisher

Walter de Gruyter GmbH

Published

2001

Language

EN

Field

Biochemistry, Genetics and Molecular Biology (Life Sciences)