Influence of glycosylation on the adsorption of Thermomyces lanuginosus lipase to hydrophobic and hydrophilic surfaces
In the pharmaceutical industry, protein drugs are modified by, for instance, glycosylation in order to obtain protein drugs with improved delivery profiles and/or increased stability. The effect of glycosylation on protein adsorption behaviour is one of the stability aspects that must be evaluated d...
Published in: | European Journal of Pharmaceutical Sciences |
---|---|
Main Authors: | , , , , , , , |
Format: | Article in Journal/Newspaper |
Language: | English |
Published: |
2010
|
Subjects: | |
Online Access: | https://research.wur.nl/en/publications/influence-of-glycosylation-on-the-adsorption-of-thermomyces-lanug https://doi.org/10.1016/j.ejps.2010.03.021 |
id |
ftunivwagenin:oai:library.wur.nl:wurpubs/407453 |
---|---|
record_format |
openpolar |
spelling |
ftunivwagenin:oai:library.wur.nl:wurpubs/407453 2024-02-04T09:54:59+01:00 Influence of glycosylation on the adsorption of Thermomyces lanuginosus lipase to hydrophobic and hydrophilic surfaces Pinholt, C. Fano, M. Wiberg, C. Hostrup, S. Bukrinsky, J.T. Frokjaer, S. Norde, W. Jorgensen, L. 2010 application/pdf https://research.wur.nl/en/publications/influence-of-glycosylation-on-the-adsorption-of-thermomyces-lanug https://doi.org/10.1016/j.ejps.2010.03.021 en eng https://edepot.wur.nl/175017 https://research.wur.nl/en/publications/influence-of-glycosylation-on-the-adsorption-of-thermomyces-lanug doi:10.1016/j.ejps.2010.03.021 info:eu-repo/semantics/restrictedAccess Wageningen University & Research European Journal of Pharmaceutical Sciences 40 (2010) 4 ISSN: 0928-0987 bacteriophage-t4 lysozyme bromide-mediated elutability candida-antarctica conformational-changes humicola-lanuginosa methylated silica surfaces protein adsorption random sequential adsorption solid-liquid interfaces tryptophan residues info:eu-repo/semantics/article Article/Letter to editor info:eu-repo/semantics/publishedVersion 2010 ftunivwagenin https://doi.org/10.1016/j.ejps.2010.03.021 2024-01-10T23:22:43Z In the pharmaceutical industry, protein drugs are modified by, for instance, glycosylation in order to obtain protein drugs with improved delivery profiles and/or increased stability. The effect of glycosylation on protein adsorption behaviour is one of the stability aspects that must be evaluated during development of glycosylated protein drug products. We have studied the effect of glycosylation on the adsorption behaviour of Thermomyces lanuginosus lipase to hydrophobic and hydrophilic surfaces using total internal reflection fluorescence, surface plasmon resonance, far-UV circular dichroism and fluorescence. Three glyco-variants were used, namely the mono-glycosylated wildtype T. lanuginosus lipase, a non-glycosylated variant and a penta-glycosylated variant, the latter two containing one and nine amino acid substitutions, respectively. All the glycosylations were N-linked and contained no charged sugar residues. Glycosylation did not affect the adsorption of wildtype T. lanuginosus lipase to the hydrophobic surfaces. The number of molecules adsorbing per unit surface area, the structural changes occurring upon adsorption, and the orientation upon adsorption were found to be unaffected by the varying glycosylation. However, the interaction with a hydrophilic surface was different between the three glyco-variants. The penta-glycosylated T. lanuginosus lipase adsorbed, in contrast to the two other glyco-variants. In conclusion, adsorption of T. lanuginosus lipase to hydrophobic surfaces was not affected by N-linked glycosylation. Only penta-glycosylated T. lanuginosus lipase adsorbed to the hydrophilic surface, apparently due to its increased net charge of +3 caused by amino acid substitutions in the primary sequence. Article in Journal/Newspaper Antarc* Antarctica Wageningen UR (University & Research Centre): Digital Library European Journal of Pharmaceutical Sciences 40 4 273 281 |
institution |
Open Polar |
collection |
Wageningen UR (University & Research Centre): Digital Library |
op_collection_id |
ftunivwagenin |
language |
English |
topic |
bacteriophage-t4 lysozyme bromide-mediated elutability candida-antarctica conformational-changes humicola-lanuginosa methylated silica surfaces protein adsorption random sequential adsorption solid-liquid interfaces tryptophan residues |
spellingShingle |
bacteriophage-t4 lysozyme bromide-mediated elutability candida-antarctica conformational-changes humicola-lanuginosa methylated silica surfaces protein adsorption random sequential adsorption solid-liquid interfaces tryptophan residues Pinholt, C. Fano, M. Wiberg, C. Hostrup, S. Bukrinsky, J.T. Frokjaer, S. Norde, W. Jorgensen, L. Influence of glycosylation on the adsorption of Thermomyces lanuginosus lipase to hydrophobic and hydrophilic surfaces |
topic_facet |
bacteriophage-t4 lysozyme bromide-mediated elutability candida-antarctica conformational-changes humicola-lanuginosa methylated silica surfaces protein adsorption random sequential adsorption solid-liquid interfaces tryptophan residues |
description |
In the pharmaceutical industry, protein drugs are modified by, for instance, glycosylation in order to obtain protein drugs with improved delivery profiles and/or increased stability. The effect of glycosylation on protein adsorption behaviour is one of the stability aspects that must be evaluated during development of glycosylated protein drug products. We have studied the effect of glycosylation on the adsorption behaviour of Thermomyces lanuginosus lipase to hydrophobic and hydrophilic surfaces using total internal reflection fluorescence, surface plasmon resonance, far-UV circular dichroism and fluorescence. Three glyco-variants were used, namely the mono-glycosylated wildtype T. lanuginosus lipase, a non-glycosylated variant and a penta-glycosylated variant, the latter two containing one and nine amino acid substitutions, respectively. All the glycosylations were N-linked and contained no charged sugar residues. Glycosylation did not affect the adsorption of wildtype T. lanuginosus lipase to the hydrophobic surfaces. The number of molecules adsorbing per unit surface area, the structural changes occurring upon adsorption, and the orientation upon adsorption were found to be unaffected by the varying glycosylation. However, the interaction with a hydrophilic surface was different between the three glyco-variants. The penta-glycosylated T. lanuginosus lipase adsorbed, in contrast to the two other glyco-variants. In conclusion, adsorption of T. lanuginosus lipase to hydrophobic surfaces was not affected by N-linked glycosylation. Only penta-glycosylated T. lanuginosus lipase adsorbed to the hydrophilic surface, apparently due to its increased net charge of +3 caused by amino acid substitutions in the primary sequence. |
format |
Article in Journal/Newspaper |
author |
Pinholt, C. Fano, M. Wiberg, C. Hostrup, S. Bukrinsky, J.T. Frokjaer, S. Norde, W. Jorgensen, L. |
author_facet |
Pinholt, C. Fano, M. Wiberg, C. Hostrup, S. Bukrinsky, J.T. Frokjaer, S. Norde, W. Jorgensen, L. |
author_sort |
Pinholt, C. |
title |
Influence of glycosylation on the adsorption of Thermomyces lanuginosus lipase to hydrophobic and hydrophilic surfaces |
title_short |
Influence of glycosylation on the adsorption of Thermomyces lanuginosus lipase to hydrophobic and hydrophilic surfaces |
title_full |
Influence of glycosylation on the adsorption of Thermomyces lanuginosus lipase to hydrophobic and hydrophilic surfaces |
title_fullStr |
Influence of glycosylation on the adsorption of Thermomyces lanuginosus lipase to hydrophobic and hydrophilic surfaces |
title_full_unstemmed |
Influence of glycosylation on the adsorption of Thermomyces lanuginosus lipase to hydrophobic and hydrophilic surfaces |
title_sort |
influence of glycosylation on the adsorption of thermomyces lanuginosus lipase to hydrophobic and hydrophilic surfaces |
publishDate |
2010 |
url |
https://research.wur.nl/en/publications/influence-of-glycosylation-on-the-adsorption-of-thermomyces-lanug https://doi.org/10.1016/j.ejps.2010.03.021 |
genre |
Antarc* Antarctica |
genre_facet |
Antarc* Antarctica |
op_source |
European Journal of Pharmaceutical Sciences 40 (2010) 4 ISSN: 0928-0987 |
op_relation |
https://edepot.wur.nl/175017 https://research.wur.nl/en/publications/influence-of-glycosylation-on-the-adsorption-of-thermomyces-lanug doi:10.1016/j.ejps.2010.03.021 |
op_rights |
info:eu-repo/semantics/restrictedAccess Wageningen University & Research |
op_doi |
https://doi.org/10.1016/j.ejps.2010.03.021 |
container_title |
European Journal of Pharmaceutical Sciences |
container_volume |
40 |
container_issue |
4 |
container_start_page |
273 |
op_container_end_page |
281 |
_version_ |
1789958829199327232 |