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Murad H, Assaad JM, Al-Shemali R, et al. Exploiting Nanobodies in the Detection and Quantification of Human Growth Hormone . Front Endocrinol (Lausanne). 2017;8:115doi: 10.3389/fendo.2017.00115.
Murad, H., Assaad, J. M., Al-Shemali, R., & Abbady, A. Q. (2017). Exploiting Nanobodies in the Detection and Quantification of Human Growth Hormone . Frontiers in endocrinology, 8115. https://doi.org/10.3389/fendo.2017.00115
Murad, Hossam, et al. "Exploiting Nanobodies in the Detection and Quantification of Human Growth Hormone ." Frontiers in endocrinology vol. 8 (2017): 115. doi: https://doi.org/10.3389/fendo.2017.00115
Murad H, Assaad JM, Al-Shemali R, Abbady AQ. Exploiting Nanobodies in the Detection and Quantification of Human Growth Hormone . Front Endocrinol (Lausanne). 2017 May 30;8:115. doi: 10.3389/fendo.2017.00115. eCollection 2017. PMID: 28611730; PMCID: PMC5447680.
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Front Endocrinol (Lausanne). 2017 May 30;8:115. doi: 10.3389/fendo.2017.00115. eCollection 2017.

Exploiting Nanobodies in the Detection and Quantification of Human Growth Hormone .

Frontiers in endocrinology

Hossam Murad, Jana Mir Assaad, Rasha Al-Shemali, Abdul Qader Abbady

Affiliations

  1. Department of Molecular Biology and Biotechnology, AECS, Damascus, Syria.

PMID: 28611730 PMCID: PMC5447680 DOI: 10.3389/fendo.2017.00115

Abstract

BACKGROUND: Monitoring blood levels of human growth hormone (hGH) in most children with short stature deficiencies is crucial for taking a decision of treatment with extended course of daily and expensive doses of recombinant hGH (rhGH or Somatropin

METHODS: A fully nanobody-based sandwich ELISA method was developed in this work for direct measurement of GH in biological samples.

RESULTS: Two major characteristics of nanobody were exploited for this goal: the robust and stable structure of the nanobody (NbGH04) used to capture hGH from tested samples, and the great ability of tailoring, enabling the display of the anti-GH detector nanobody (NbGH07) on the tip of M13-phage. Such huge, stable, and easy-to-prepare phage-Nb was used in ELISA to provide an amplified signal. Previously, NbGH04 was retrieved on immobilized hGH by phage display from a wide "immune" cDNA library prepared from a hGH-immunized camel. Here, and in order to assure epitope heterogeneity, NbGH07 was isolated from the same library using NbGH04-captured hGH as bait. Interaction of both nanobodies with hGH was characterized and compared with different anti-GH nanobodies and antibodies. The sensitivity (~0.5 ng/ml) and stability of the nanobody-base sandwich ELISA were assessed using rhGH before testing in the quantification of hGH in blood sera and cell culture supernatants.

CONCLUSION: In regard to all advantages of nanobodies; stability, solubility, production affordability in

Keywords: VHH; biotinylation; camel; doping detection; growth hormone; nanobody; phage display; recombinant antibody

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