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Issue 10, 2009
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Surface plasmon resonance biosensor for direct detection of antibodies against human growth hormone

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Abstract

A direct, label-free detection method of antibodies against the human growth hormone (anti-HGH) using a surface plasmon resonance (SPR) biosensor is reported. The sensing surface of the surface plasmon resonance biosensor chip (SPR-chip) was modified by covalent coupling of the human growth hormone (HGH) to the self-assembled monolayer of 11-mercaptoundecanoic acid (MUA). HGH was immobilized viaprimary amine groups after activation of the MUAcarboxyl groups with a mixture of N-hydroxysuccinimide (NHS), and N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (EDC). The specific binding of monoclonal anti-HGH antibody on the HGH-modified surface was examined in the concentration range from 0.25 nM to 10 µM. The experimentally observed detection minimum for anti-HGH was 2.47 nM. A single immunoassay cycle could be done within 30 min including the HGH and anti-HGH association, HGH/anti-HGH complex dissociation and surface regeneration steps. The SPR biosensor response for repeatable detections of anti-HGH was highly reproducible and very stable. On the SPR-chip the formed HGH and anti-HGH complex (HGH/anti-HGH) could be gently dissociated and the sensing surface might be regenerated by 50 mM NaOH and 0.5% sodium dodecylsulfate (SDS) solution. Any changes in the original baseline level were detected during the 40 detection–regeneration cycles. This means that damage of the immobilized HGH-based sensitive layer during regeneration was minimal. It was demonstrated that the developed SPR-chip could be stored for at least 21 days before use without considerable loss of sensitivity towards anti-HGH.

Graphical abstract: Surface plasmon resonance biosensor for direct detection of antibodies against human growth hormone

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Publication details

The article was received on 09 Apr 2009, accepted on 17 Jul 2009 and first published on 31 Jul 2009


Article type: Paper
DOI: 10.1039/B907315A
Citation: Analyst, 2009,134, 2051-2057
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    Surface plasmon resonance biosensor for direct detection of antibodies against human growth hormone

    A. Kausaite-Minkstimiene, A. Ramanaviciene and A. Ramanavicius, Analyst, 2009, 134, 2051
    DOI: 10.1039/B907315A

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