Increased adsorption of histidine-tagged proteins onto tissue culture polystyrene
Research output: Contribution to journal › Journal article › Research › peer-review
Standard
Increased adsorption of histidine-tagged proteins onto tissue culture polystyrene. / Holmberg, Maria; Hansen, Thomas Steen; Lind, Johan Ulrik; Hjortø, Gertrud Malene.
In: Colloids and Surfaces B: Biointerfaces, Vol. 92, 01.04.2012, p. 286-292.Research output: Contribution to journal › Journal article › Research › peer-review
Harvard
APA
Vancouver
Author
Bibtex
}
RIS
TY - JOUR
T1 - Increased adsorption of histidine-tagged proteins onto tissue culture polystyrene
AU - Holmberg, Maria
AU - Hansen, Thomas Steen
AU - Lind, Johan Ulrik
AU - Hjortø, Gertrud Malene
N1 - Copyright © 2011 Elsevier B.V. All rights reserved.
PY - 2012/4/1
Y1 - 2012/4/1
N2 - In this study we compare histidine-tagged and native proteins with regards to adsorption properties. We observe significantly increased adsorption of proteins with an incorporated polyhistidine amino acid motif (HIS-tag) onto tissue culture polystyrene (TCPS) compared to similar proteins without a HIS-tag. The effect is not observed on polystyrene (PS). Adsorption experiments have been performed at physiological pH (7.4) and the effect was only observed for the investigated proteins that have pI values below or around 7.4. Competitive adsorption experiments with imidazole and ethylenediaminetetraacetic acid (EDTA), as well as adsorption performed at different pH and ionic strength indicates that the high adsorption is caused by electrostatic interaction between negatively charged carboxylate groups on the TCPS surface and positively charged histidine residues in the proteins. Pre-adsorption of bovine serum albumin (BSA) does not decrease the adsorption of HIS-tagged proteins onto TCPS. Our findings identify a potential problem in using HIS-tagged signalling molecule in assays with cells cultured on TCPS, since the concentration of the molecule in solution might be affected and this could critically influence the assay outcome.
AB - In this study we compare histidine-tagged and native proteins with regards to adsorption properties. We observe significantly increased adsorption of proteins with an incorporated polyhistidine amino acid motif (HIS-tag) onto tissue culture polystyrene (TCPS) compared to similar proteins without a HIS-tag. The effect is not observed on polystyrene (PS). Adsorption experiments have been performed at physiological pH (7.4) and the effect was only observed for the investigated proteins that have pI values below or around 7.4. Competitive adsorption experiments with imidazole and ethylenediaminetetraacetic acid (EDTA), as well as adsorption performed at different pH and ionic strength indicates that the high adsorption is caused by electrostatic interaction between negatively charged carboxylate groups on the TCPS surface and positively charged histidine residues in the proteins. Pre-adsorption of bovine serum albumin (BSA) does not decrease the adsorption of HIS-tagged proteins onto TCPS. Our findings identify a potential problem in using HIS-tagged signalling molecule in assays with cells cultured on TCPS, since the concentration of the molecule in solution might be affected and this could critically influence the assay outcome.
KW - Adsorption
KW - Animals
KW - Histidine
KW - Humans
KW - Hydrogen-Ion Concentration
KW - Polystyrenes
KW - Recombinant Fusion Proteins
KW - Tissue Culture Techniques
KW - Journal Article
U2 - 10.1016/j.colsurfb.2011.12.001
DO - 10.1016/j.colsurfb.2011.12.001
M3 - Journal article
C2 - 22225945
VL - 92
SP - 286
EP - 292
JO - Colloids and Surfaces B: Biointerfaces
JF - Colloids and Surfaces B: Biointerfaces
SN - 0927-7765
ER -
ID: 182199270