Surface plasmon resonance biosensor for direct detection of antibodies against human growth hormone
- 31 July 2009
- journal article
- research article
- Published by Royal Society of Chemistry (RSC) in The Analyst
- Vol. 134 (10), 2051-2057
- https://doi.org/10.1039/b907315a
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.Keywords
This publication has 41 references indexed in Scilit:
- Growth hormone excess and the development of growth hormone receptor antagonistsExperimental Physiology, 2008
- Which children with idiopathic short stature should receive growth hormone therapy?Nature Clinical Practice Endocrinology & Metabolism, 2007
- Growth Hormone Therapy for Short Children Born Small for Gestational AgeHormone Research in Paediatrics, 2007
- Growth Hormone Treatment of Early Growth Failure in Toddlers with Turner Syndrome: A Randomized, Controlled, Multicenter TrialJournal of Clinical Endocrinology & Metabolism, 2007
- Two years of growth hormone therapy in young children with Prader–Willi syndrome: Physical and neurodevelopmental benefitsAmerican Journal of Medical Genetics Part A, 2006
- Evaluation and Treatment of Adult Growth Hormone Deficiency: An Endocrine Society Clinical Practice GuidelineJournal of Clinical Endocrinology & Metabolism, 2006
- Growth hormone for children with chronic renal failure and on dialysisPediatric Nephrology, 2005
- Growth Hormone Antibodies Formation in Patients Treated with Recombinant Human Growth HormoneInternational Journal of Immunopathology and Pharmacology, 2004
- Construction of a specific and sensitive sandwich enzyme immunoassay for 20 kDa human growth hormoneJournal of Immunological Methods, 1998
- Development of Anti‐hGH Antibodies during Therapy with Authentic Human Growth HormoneActa Paediatrica, 1991