Gravimetric antigen detection utilizing antibody-modified lipid bilayers
(2005) In Analytical Biochemistry 345(1). p.72-80- Abstract
- Lipid bilayers containing 5% nitrilotriacetic acid (NTA) lipids supported on SiO2 have been used as a template for immobilization of oligohistidine-tagged single-chained antibody fragments (scFvs) directed against cholera toxin. It was demonstrated that histidine-tagged scFvs could be equally efficiently coupled to an NTA-Ni2+-containing lipid bilayer from a purified sample as from an expression supernatant, thereby providing a coupling method that eliminates time-consuming protein prepurification steps. Irrespective of whether the coupling was made from the unpurified or purified antibody preparation, the template proved to be efficient for antigen (cholera toxin) detection, verified using quartz crystal microbalance with dissipation... (More)
- Lipid bilayers containing 5% nitrilotriacetic acid (NTA) lipids supported on SiO2 have been used as a template for immobilization of oligohistidine-tagged single-chained antibody fragments (scFvs) directed against cholera toxin. It was demonstrated that histidine-tagged scFvs could be equally efficiently coupled to an NTA-Ni2+-containing lipid bilayer from a purified sample as from an expression supernatant, thereby providing a coupling method that eliminates time-consuming protein prepurification steps. Irrespective of whether the coupling was made from the unpurified or purified antibody preparation, the template proved to be efficient for antigen (cholera toxin) detection, verified using quartz crystal microbalance with dissipation monitoring. In addition, via a secondary amplification step using lipid vesicles containing GM, (the natural membrane receptor for cholera toxin), the detection limit of cholera toxin was less than 750 pM. To further strengthen the coupling of scFvs to the lipid bilayer, scFvs containing two histidine tags, instead of just one tag, were also evaluated. The increased coupling strength provided via the bivalent anchoring significantly reduced scFv displacement in complex solutions containing large amounts of histidine-containing proteins, verified via cholera toxin detection in serum. (C) 2005 Elsevier Inc. All rights reserved. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/152123
- author
- Larsson, C ; Bramfeldt, H ; Wingren, Christer LU ; Borrebaeck, Carl LU and Hook, F
- organization
- publishing date
- 2005
- type
- Contribution to journal
- publication status
- published
- subject
- in
- Analytical Biochemistry
- volume
- 345
- issue
- 1
- pages
- 72 - 80
- publisher
- Elsevier
- external identifiers
-
- wos:000232431700009
- pmid:16139234
- scopus:27644563072
- pmid:16139234
- ISSN
- 1096-0309
- DOI
- 10.1016/j.ab.2005.05.031
- language
- English
- LU publication?
- yes
- id
- 060ebadb-2968-4f89-b9cf-039049829ddd (old id 152123)
- date added to LUP
- 2016-04-01 11:56:21
- date last changed
- 2022-01-26 20:27:16
@article{060ebadb-2968-4f89-b9cf-039049829ddd,
abstract = {{Lipid bilayers containing 5% nitrilotriacetic acid (NTA) lipids supported on SiO2 have been used as a template for immobilization of oligohistidine-tagged single-chained antibody fragments (scFvs) directed against cholera toxin. It was demonstrated that histidine-tagged scFvs could be equally efficiently coupled to an NTA-Ni2+-containing lipid bilayer from a purified sample as from an expression supernatant, thereby providing a coupling method that eliminates time-consuming protein prepurification steps. Irrespective of whether the coupling was made from the unpurified or purified antibody preparation, the template proved to be efficient for antigen (cholera toxin) detection, verified using quartz crystal microbalance with dissipation monitoring. In addition, via a secondary amplification step using lipid vesicles containing GM, (the natural membrane receptor for cholera toxin), the detection limit of cholera toxin was less than 750 pM. To further strengthen the coupling of scFvs to the lipid bilayer, scFvs containing two histidine tags, instead of just one tag, were also evaluated. The increased coupling strength provided via the bivalent anchoring significantly reduced scFv displacement in complex solutions containing large amounts of histidine-containing proteins, verified via cholera toxin detection in serum. (C) 2005 Elsevier Inc. All rights reserved.}},
author = {{Larsson, C and Bramfeldt, H and Wingren, Christer and Borrebaeck, Carl and Hook, F}},
issn = {{1096-0309}},
language = {{eng}},
number = {{1}},
pages = {{72--80}},
publisher = {{Elsevier}},
series = {{Analytical Biochemistry}},
title = {{Gravimetric antigen detection utilizing antibody-modified lipid bilayers}},
url = {{http://dx.doi.org/10.1016/j.ab.2005.05.031}},
doi = {{10.1016/j.ab.2005.05.031}},
volume = {{345}},
year = {{2005}},
}