Cargando…

Semisynthetic Bioluminescent Sensor Proteins for Direct Detection of Antibodies and Small Molecules in Solution

[Image: see text] Single-step immunoassays that can be performed directly in solution are ideally suited for point-of-care diagnostics. Our group recently developed a new platform of bioluminescent sensor proteins (LUMABS; LUMinescent AntiBody Sensor) that allow antibody detection in blood plasma. T...

Descripción completa

Detalles Bibliográficos
Autores principales: Arts, Remco, Ludwig, Susann K. J., van Gerven, Benice C. B., Estirado, Eva Magdalena, Milroy, Lech-Gustav, Merkx, Maarten
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Chemical Society 2017
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5706068/
https://www.ncbi.nlm.nih.gov/pubmed/29037030
http://dx.doi.org/10.1021/acssensors.7b00695
_version_ 1783282155184455680
author Arts, Remco
Ludwig, Susann K. J.
van Gerven, Benice C. B.
Estirado, Eva Magdalena
Milroy, Lech-Gustav
Merkx, Maarten
author_facet Arts, Remco
Ludwig, Susann K. J.
van Gerven, Benice C. B.
Estirado, Eva Magdalena
Milroy, Lech-Gustav
Merkx, Maarten
author_sort Arts, Remco
collection PubMed
description [Image: see text] Single-step immunoassays that can be performed directly in solution are ideally suited for point-of-care diagnostics. Our group recently developed a new platform of bioluminescent sensor proteins (LUMABS; LUMinescent AntiBody Sensor) that allow antibody detection in blood plasma. Thus far, LUMABS has been limited to the detection of antibodies recognizing natural peptide epitopes. Here, we report the development of semisynthetic LUMABS sensors that recognize nonpeptide epitopes. The non-natural amino acid para-azidophenylalanine was introduced at the position of the original antibody-recognition sites as a chemical handle to enable site-specific conjugation of synthetic epitope molecules coupled to a dibenzocylcooctyne moiety via strain-promoted click chemistry. The approach was successfully demonstrated by developing semisynthetic LUMABS sensors for antibodies targeting the small molecules dinitrophenol and creatinine (DNP-LUMABS and CR-LUMABS) with affinities of 5.8 pM and 1.3 nM, respectively. An important application of these semisynthetic LUMABS is the detection of small molecules using a competitive assay format, which is demonstrated here for the detection of creatinine. Using a preassembled complex of CR-LUMABS and an anti-creatinine antibody, the detection of high micromolar concentrations of creatinine was possible both in buffer and in 1:1 diluted blood plasma. The use of semisynthetic LUMABS sensors significantly expands the range of antibody targets and enables the application of LUMABS sensors for the ratiometric bioluminescent detection of small molecules using a competitive immunoassay format.
format Online
Article
Text
id pubmed-5706068
institution National Center for Biotechnology Information
language English
publishDate 2017
publisher American Chemical Society
record_format MEDLINE/PubMed
spelling pubmed-57060682017-11-30 Semisynthetic Bioluminescent Sensor Proteins for Direct Detection of Antibodies and Small Molecules in Solution Arts, Remco Ludwig, Susann K. J. van Gerven, Benice C. B. Estirado, Eva Magdalena Milroy, Lech-Gustav Merkx, Maarten ACS Sens [Image: see text] Single-step immunoassays that can be performed directly in solution are ideally suited for point-of-care diagnostics. Our group recently developed a new platform of bioluminescent sensor proteins (LUMABS; LUMinescent AntiBody Sensor) that allow antibody detection in blood plasma. Thus far, LUMABS has been limited to the detection of antibodies recognizing natural peptide epitopes. Here, we report the development of semisynthetic LUMABS sensors that recognize nonpeptide epitopes. The non-natural amino acid para-azidophenylalanine was introduced at the position of the original antibody-recognition sites as a chemical handle to enable site-specific conjugation of synthetic epitope molecules coupled to a dibenzocylcooctyne moiety via strain-promoted click chemistry. The approach was successfully demonstrated by developing semisynthetic LUMABS sensors for antibodies targeting the small molecules dinitrophenol and creatinine (DNP-LUMABS and CR-LUMABS) with affinities of 5.8 pM and 1.3 nM, respectively. An important application of these semisynthetic LUMABS is the detection of small molecules using a competitive assay format, which is demonstrated here for the detection of creatinine. Using a preassembled complex of CR-LUMABS and an anti-creatinine antibody, the detection of high micromolar concentrations of creatinine was possible both in buffer and in 1:1 diluted blood plasma. The use of semisynthetic LUMABS sensors significantly expands the range of antibody targets and enables the application of LUMABS sensors for the ratiometric bioluminescent detection of small molecules using a competitive immunoassay format. American Chemical Society 2017-10-17 2017-11-22 /pmc/articles/PMC5706068/ /pubmed/29037030 http://dx.doi.org/10.1021/acssensors.7b00695 Text en Copyright © 2017 American Chemical Society This is an open access article published under a Creative Commons Non-Commercial No Derivative Works (CC-BY-NC-ND) Attribution License (http://pubs.acs.org/page/policy/authorchoice_ccbyncnd_termsofuse.html) , which permits copying and redistribution of the article, and creation of adaptations, all for non-commercial purposes.
spellingShingle Arts, Remco
Ludwig, Susann K. J.
van Gerven, Benice C. B.
Estirado, Eva Magdalena
Milroy, Lech-Gustav
Merkx, Maarten
Semisynthetic Bioluminescent Sensor Proteins for Direct Detection of Antibodies and Small Molecules in Solution
title Semisynthetic Bioluminescent Sensor Proteins for Direct Detection of Antibodies and Small Molecules in Solution
title_full Semisynthetic Bioluminescent Sensor Proteins for Direct Detection of Antibodies and Small Molecules in Solution
title_fullStr Semisynthetic Bioluminescent Sensor Proteins for Direct Detection of Antibodies and Small Molecules in Solution
title_full_unstemmed Semisynthetic Bioluminescent Sensor Proteins for Direct Detection of Antibodies and Small Molecules in Solution
title_short Semisynthetic Bioluminescent Sensor Proteins for Direct Detection of Antibodies and Small Molecules in Solution
title_sort semisynthetic bioluminescent sensor proteins for direct detection of antibodies and small molecules in solution
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5706068/
https://www.ncbi.nlm.nih.gov/pubmed/29037030
http://dx.doi.org/10.1021/acssensors.7b00695
work_keys_str_mv AT artsremco semisyntheticbioluminescentsensorproteinsfordirectdetectionofantibodiesandsmallmoleculesinsolution
AT ludwigsusannkj semisyntheticbioluminescentsensorproteinsfordirectdetectionofantibodiesandsmallmoleculesinsolution
AT vangervenbenicecb semisyntheticbioluminescentsensorproteinsfordirectdetectionofantibodiesandsmallmoleculesinsolution
AT estiradoevamagdalena semisyntheticbioluminescentsensorproteinsfordirectdetectionofantibodiesandsmallmoleculesinsolution
AT milroylechgustav semisyntheticbioluminescentsensorproteinsfordirectdetectionofantibodiesandsmallmoleculesinsolution
AT merkxmaarten semisyntheticbioluminescentsensorproteinsfordirectdetectionofantibodiesandsmallmoleculesinsolution