Molecular and functional characterisation of a fusion protein suited for tumour specific prodrug activation.

A fusion protein consisting of the humanised Fab fragment of the anti CEA MAb BW 431 and the human beta-glucuronidase was expressed in BHK cells. Functional testing revealed that the specificity and avidity of the humanised V region was similar to the original murine MAb BW 431. Furthermore, the enz...

Descripción completa

Detalles Bibliográficos
Autores principales: Bosslet, K., Czech, J., Lorenz, P., Sedlacek, H. H., Schuermann, M., Seemann, G.
Formato: Texto
Lenguaje:English
Publicado: Nature Publishing Group 1992
Materias:
Research Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1977731/
https://www.ncbi.nlm.nih.gov/pubmed/1739623
_version_ 1782135325176365056
author Bosslet, K.
Czech, J.
Lorenz, P.
Sedlacek, H. H.
Schuermann, M.
Seemann, G.
author_facet Bosslet, K.
Czech, J.
Lorenz, P.
Sedlacek, H. H.
Schuermann, M.
Seemann, G.
author_sort Bosslet, K.
collection PubMed
description A fusion protein consisting of the humanised Fab fragment of the anti CEA MAb BW 431 and the human beta-glucuronidase was expressed in BHK cells. Functional testing revealed that the specificity and avidity of the humanised V region was similar to the original murine MAb BW 431. Furthermore, the enzymatic activity, pH sensitivity and stability of the human beta-glucuronidase in the fusion protein was comparable to the activity of recombinant human beta-glucuronidase. Using anti-idiotype affinity chromatography, two molecules of a molecular weight of 125 kDa or 250 kDa could be visualized under nonreducing conditions in SDS-PAGE. Reducing conditions revealed a 25 kDa light and 100 kDa heavy chain. Due to its suitable biological characteristics this fusion protein might be an appropriate molecule allowing a site specific antibody directed enzyme prodrug therapy (ADEPT) in vivo. IMAGES:
format Text
id pubmed-1977731
institution National Center for Biotechnology Information
language English
publishDate 1992
publisher Nature Publishing Group
record_format MEDLINE/PubMed
spelling pubmed-19777312009-09-10 Molecular and functional characterisation of a fusion protein suited for tumour specific prodrug activation. Bosslet, K. Czech, J. Lorenz, P. Sedlacek, H. H. Schuermann, M. Seemann, G. Br J Cancer Research Article A fusion protein consisting of the humanised Fab fragment of the anti CEA MAb BW 431 and the human beta-glucuronidase was expressed in BHK cells. Functional testing revealed that the specificity and avidity of the humanised V region was similar to the original murine MAb BW 431. Furthermore, the enzymatic activity, pH sensitivity and stability of the human beta-glucuronidase in the fusion protein was comparable to the activity of recombinant human beta-glucuronidase. Using anti-idiotype affinity chromatography, two molecules of a molecular weight of 125 kDa or 250 kDa could be visualized under nonreducing conditions in SDS-PAGE. Reducing conditions revealed a 25 kDa light and 100 kDa heavy chain. Due to its suitable biological characteristics this fusion protein might be an appropriate molecule allowing a site specific antibody directed enzyme prodrug therapy (ADEPT) in vivo. IMAGES: Nature Publishing Group 1992-02 /pmc/articles/PMC1977731/ /pubmed/1739623 Text en https://creativecommons.org/licenses/by/4.0/This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit https://creativecommons.org/licenses/by/4.0/.
spellingShingle Research Article
Bosslet, K.
Czech, J.
Lorenz, P.
Sedlacek, H. H.
Schuermann, M.
Seemann, G.
Molecular and functional characterisation of a fusion protein suited for tumour specific prodrug activation.
title Molecular and functional characterisation of a fusion protein suited for tumour specific prodrug activation.
title_full Molecular and functional characterisation of a fusion protein suited for tumour specific prodrug activation.
title_fullStr Molecular and functional characterisation of a fusion protein suited for tumour specific prodrug activation.
title_full_unstemmed Molecular and functional characterisation of a fusion protein suited for tumour specific prodrug activation.
title_short Molecular and functional characterisation of a fusion protein suited for tumour specific prodrug activation.
title_sort molecular and functional characterisation of a fusion protein suited for tumour specific prodrug activation.
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1977731/
https://www.ncbi.nlm.nih.gov/pubmed/1739623
work_keys_str_mv AT bossletk molecularandfunctionalcharacterisationofafusionproteinsuitedfortumourspecificprodrugactivation
AT czechj molecularandfunctionalcharacterisationofafusionproteinsuitedfortumourspecificprodrugactivation
AT lorenzp molecularandfunctionalcharacterisationofafusionproteinsuitedfortumourspecificprodrugactivation
AT sedlacekhh molecularandfunctionalcharacterisationofafusionproteinsuitedfortumourspecificprodrugactivation
AT schuermannm molecularandfunctionalcharacterisationofafusionproteinsuitedfortumourspecificprodrugactivation
AT seemanng molecularandfunctionalcharacterisationofafusionproteinsuitedfortumourspecificprodrugactivation

Ejemplares similares