Cargando…

Microsphere Peptide-Based Immunoassay for the Detection of Recombinant Bovine Somatotropin in Injection Preparations

The use of peptides in immunoassays can be favored over the use of the full protein when more cost effective or less toxic approaches are needed, or when access to the full protein is lacking. Due to restricted access to recombinant bovine somatotropin (rbST), a protein enhancing growth and lactatin...

Descripción completa

Detalles Bibliográficos
Autores principales: Smits, Nathalie G. E., Bovee, Toine F. H., Pujari, Sidharam P., van Ginkel, Leendert A., Nielen, Michel W. F., Albada, Bauke
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8946150/
https://www.ncbi.nlm.nih.gov/pubmed/35323408
http://dx.doi.org/10.3390/bios12030138
_version_ 1784674126103838720
author Smits, Nathalie G. E.
Bovee, Toine F. H.
Pujari, Sidharam P.
van Ginkel, Leendert A.
Nielen, Michel W. F.
Albada, Bauke
author_facet Smits, Nathalie G. E.
Bovee, Toine F. H.
Pujari, Sidharam P.
van Ginkel, Leendert A.
Nielen, Michel W. F.
Albada, Bauke
author_sort Smits, Nathalie G. E.
collection PubMed
description The use of peptides in immunoassays can be favored over the use of the full protein when more cost effective or less toxic approaches are needed, or when access to the full protein is lacking. Due to restricted access to recombinant bovine somatotropin (rbST), a protein enhancing growth and lactating performances of livestock, which use has been banned in the EU, Canada and Australia (amongst others), we developed a peptide-based biorecognition assay on an imaging planar array analyzer. For this, we identified the rbST epitope that is responsible for binding to the rbST-targeting monoclonal antibody 4H12 (MAb 4H12) to be (115)DLEEGILALMR(125). This linear peptide was synthesized and coupled to microspheres, after which it was tested in a biorecognition competitive inhibition assay format. We observed IC(50) values of approximately 0.11 μg mL(−)(1), which are lower than observed for the full rbST protein (IC(50) = 0.20 μg mL(−1)). Importantly, there was no binding with the scrambled peptide. Preliminary results of directly coupled peptides in a microsphere biorecognition assay for detection of rbST are presented. Real-life applicability for detection of somatotropins (STs) in injection preparations of bovine-, porcine- and equine ST are shown. This newly developed immunoassay strongly supports future developments of peptide-based immunoassays to circumvent the limited access to the full protein.
format Online
Article
Text
id pubmed-8946150
institution National Center for Biotechnology Information
language English
publishDate 2022
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-89461502022-03-25 Microsphere Peptide-Based Immunoassay for the Detection of Recombinant Bovine Somatotropin in Injection Preparations Smits, Nathalie G. E. Bovee, Toine F. H. Pujari, Sidharam P. van Ginkel, Leendert A. Nielen, Michel W. F. Albada, Bauke Biosensors (Basel) Article The use of peptides in immunoassays can be favored over the use of the full protein when more cost effective or less toxic approaches are needed, or when access to the full protein is lacking. Due to restricted access to recombinant bovine somatotropin (rbST), a protein enhancing growth and lactating performances of livestock, which use has been banned in the EU, Canada and Australia (amongst others), we developed a peptide-based biorecognition assay on an imaging planar array analyzer. For this, we identified the rbST epitope that is responsible for binding to the rbST-targeting monoclonal antibody 4H12 (MAb 4H12) to be (115)DLEEGILALMR(125). This linear peptide was synthesized and coupled to microspheres, after which it was tested in a biorecognition competitive inhibition assay format. We observed IC(50) values of approximately 0.11 μg mL(−)(1), which are lower than observed for the full rbST protein (IC(50) = 0.20 μg mL(−1)). Importantly, there was no binding with the scrambled peptide. Preliminary results of directly coupled peptides in a microsphere biorecognition assay for detection of rbST are presented. Real-life applicability for detection of somatotropins (STs) in injection preparations of bovine-, porcine- and equine ST are shown. This newly developed immunoassay strongly supports future developments of peptide-based immunoassays to circumvent the limited access to the full protein. MDPI 2022-02-22 /pmc/articles/PMC8946150/ /pubmed/35323408 http://dx.doi.org/10.3390/bios12030138 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Smits, Nathalie G. E.
Bovee, Toine F. H.
Pujari, Sidharam P.
van Ginkel, Leendert A.
Nielen, Michel W. F.
Albada, Bauke
Microsphere Peptide-Based Immunoassay for the Detection of Recombinant Bovine Somatotropin in Injection Preparations
title Microsphere Peptide-Based Immunoassay for the Detection of Recombinant Bovine Somatotropin in Injection Preparations
title_full Microsphere Peptide-Based Immunoassay for the Detection of Recombinant Bovine Somatotropin in Injection Preparations
title_fullStr Microsphere Peptide-Based Immunoassay for the Detection of Recombinant Bovine Somatotropin in Injection Preparations
title_full_unstemmed Microsphere Peptide-Based Immunoassay for the Detection of Recombinant Bovine Somatotropin in Injection Preparations
title_short Microsphere Peptide-Based Immunoassay for the Detection of Recombinant Bovine Somatotropin in Injection Preparations
title_sort microsphere peptide-based immunoassay for the detection of recombinant bovine somatotropin in injection preparations
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8946150/
https://www.ncbi.nlm.nih.gov/pubmed/35323408
http://dx.doi.org/10.3390/bios12030138
work_keys_str_mv AT smitsnathaliege microspherepeptidebasedimmunoassayforthedetectionofrecombinantbovinesomatotropinininjectionpreparations
AT boveetoinefh microspherepeptidebasedimmunoassayforthedetectionofrecombinantbovinesomatotropinininjectionpreparations
AT pujarisidharamp microspherepeptidebasedimmunoassayforthedetectionofrecombinantbovinesomatotropinininjectionpreparations
AT vanginkelleenderta microspherepeptidebasedimmunoassayforthedetectionofrecombinantbovinesomatotropinininjectionpreparations
AT nielenmichelwf microspherepeptidebasedimmunoassayforthedetectionofrecombinantbovinesomatotropinininjectionpreparations
AT albadabauke microspherepeptidebasedimmunoassayforthedetectionofrecombinantbovinesomatotropinininjectionpreparations