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Issue 7, 2013
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Site-directed antibody immobilization using a protein A–gold binding domain fusion protein for enhanced SPR immunosensing

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Abstract

We have implemented a novel strategy for the oriented immobilization of antibodies onto a gold surface based on the use of a fusion protein, the protein A–gold binding domain (PAG). PAG consists of a gold binding peptide (GBP) coupled to the immunoglobulin-binding domains of staphylococcal protein A. This fusion protein provides an easy and fast oriented immobilization of antibodies preserving its native structure, while leaving the antigen binding sites (Fab) freely exposed. Using this immobilization strategy, we have demonstrated the performance of the immunosensing of the human Growth Hormone by SPR. A limit of detection of 90 ng mL−1 was obtained with an inter-chip variability lower than 7%. The comparison of this method with other strategies for the direct immobilization of antibodies over gold surfaces has showed the enhanced sensitivity provided by the PAG approach.

Graphical abstract: Site-directed antibody immobilization using a protein A–gold binding domain fusion protein for enhanced SPR immunosensing

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Publication details

The article was received on 16 Oct 2012, accepted on 22 Jan 2013 and first published on 23 Jan 2013


Article type: Paper
DOI: 10.1039/C3AN36498D
Analyst, 2013,138, 2023-2031

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    Site-directed antibody immobilization using a protein A–gold binding domain fusion protein for enhanced SPR immunosensing

    E. de Juan-Franco, A. Caruz, J. R. Pedrajas and L. M. Lechuga, Analyst, 2013, 138, 2023
    DOI: 10.1039/C3AN36498D

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