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 |
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2010
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ftunigroningenpu:oai:pure.rug.nl:publications/5fe66177-aa41-47b2-9c48-e1c45ee175cd 2024-06-02T07:56:00+00:00 Influence of glycosylation on the adsorption of Thermomyces lanuginosus lipase to hydrophobic and hydrophilic surfaces Pinholt, Charlotte Fano, Mathias Wiberg, Charlotte Hostrup, Susanne Bukrinsky, Jens Thostrup Frokjaer, Sven Norde, Willem Jorgensen, Lene 2010-07-11 https://hdl.handle.net/11370/5fe66177-aa41-47b2-9c48-e1c45ee175cd https://research.rug.nl/en/publications/5fe66177-aa41-47b2-9c48-e1c45ee175cd https://doi.org/10.1016/j.ejps.2010.03.021 eng eng https://research.rug.nl/en/publications/5fe66177-aa41-47b2-9c48-e1c45ee175cd info:eu-repo/semantics/restrictedAccess Pinholt , C , Fano , M , Wiberg , C , Hostrup , S , Bukrinsky , J T , Frokjaer , S , Norde , W & Jorgensen , L 2010 , ' Influence of glycosylation on the adsorption of Thermomyces lanuginosus lipase to hydrophobic and hydrophilic surfaces ' , European Journal of Pharmaceutical Sciences , vol. 40 , no. 4 , pp. 273-281 . https://doi.org/10.1016/j.ejps.2010.03.021 Glycosylation Protein adsorption TIRF CD SPR Thermomyces lanuginosus lipase RANDOM SEQUENTIAL ADSORPTION BROMIDE-MEDIATED ELUTABILITY METHYLATED SILICA SURFACES SOLID-LIQUID INTERFACES HUMICOLA-LANUGINOSA BACTERIOPHAGE-T4 LYSOZYME CONFORMATIONAL-CHANGES TRYPTOPHAN RESIDUES CANDIDA-ANTARCTICA article 2010 ftunigroningenpu https://doi.org/10.1016/j.ejps.2010.03.021 2024-05-07T18:43:19Z 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. (C) 2010 Elsevier BM. All rights reserved. Article in Journal/Newspaper Antarc* Antarctica University of Groningen research database European Journal of Pharmaceutical Sciences 40 4 273 281 |
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Open Polar |
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University of Groningen research database |
op_collection_id |
ftunigroningenpu |
language |
English |
topic |
Glycosylation Protein adsorption TIRF CD SPR Thermomyces lanuginosus lipase RANDOM SEQUENTIAL ADSORPTION BROMIDE-MEDIATED ELUTABILITY METHYLATED SILICA SURFACES SOLID-LIQUID INTERFACES HUMICOLA-LANUGINOSA BACTERIOPHAGE-T4 LYSOZYME CONFORMATIONAL-CHANGES TRYPTOPHAN RESIDUES CANDIDA-ANTARCTICA |
spellingShingle |
Glycosylation Protein adsorption TIRF CD SPR Thermomyces lanuginosus lipase RANDOM SEQUENTIAL ADSORPTION BROMIDE-MEDIATED ELUTABILITY METHYLATED SILICA SURFACES SOLID-LIQUID INTERFACES HUMICOLA-LANUGINOSA BACTERIOPHAGE-T4 LYSOZYME CONFORMATIONAL-CHANGES TRYPTOPHAN RESIDUES CANDIDA-ANTARCTICA Pinholt, Charlotte Fano, Mathias Wiberg, Charlotte Hostrup, Susanne Bukrinsky, Jens Thostrup Frokjaer, Sven Norde, Willem Jorgensen, Lene Influence of glycosylation on the adsorption of Thermomyces lanuginosus lipase to hydrophobic and hydrophilic surfaces |
topic_facet |
Glycosylation Protein adsorption TIRF CD SPR Thermomyces lanuginosus lipase RANDOM SEQUENTIAL ADSORPTION BROMIDE-MEDIATED ELUTABILITY METHYLATED SILICA SURFACES SOLID-LIQUID INTERFACES HUMICOLA-LANUGINOSA BACTERIOPHAGE-T4 LYSOZYME CONFORMATIONAL-CHANGES TRYPTOPHAN RESIDUES CANDIDA-ANTARCTICA |
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. (C) 2010 Elsevier BM. All rights reserved. |
format |
Article in Journal/Newspaper |
author |
Pinholt, Charlotte Fano, Mathias Wiberg, Charlotte Hostrup, Susanne Bukrinsky, Jens Thostrup Frokjaer, Sven Norde, Willem Jorgensen, Lene |
author_facet |
Pinholt, Charlotte Fano, Mathias Wiberg, Charlotte Hostrup, Susanne Bukrinsky, Jens Thostrup Frokjaer, Sven Norde, Willem Jorgensen, Lene |
author_sort |
Pinholt, Charlotte |
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://hdl.handle.net/11370/5fe66177-aa41-47b2-9c48-e1c45ee175cd https://research.rug.nl/en/publications/5fe66177-aa41-47b2-9c48-e1c45ee175cd https://doi.org/10.1016/j.ejps.2010.03.021 |
genre |
Antarc* Antarctica |
genre_facet |
Antarc* Antarctica |
op_source |
Pinholt , C , Fano , M , Wiberg , C , Hostrup , S , Bukrinsky , J T , Frokjaer , S , Norde , W & Jorgensen , L 2010 , ' Influence of glycosylation on the adsorption of Thermomyces lanuginosus lipase to hydrophobic and hydrophilic surfaces ' , European Journal of Pharmaceutical Sciences , vol. 40 , no. 4 , pp. 273-281 . https://doi.org/10.1016/j.ejps.2010.03.021 |
op_relation |
https://research.rug.nl/en/publications/5fe66177-aa41-47b2-9c48-e1c45ee175cd |
op_rights |
info:eu-repo/semantics/restrictedAccess |
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 |
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281 |
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1800752647965245440 |