by Marciello, Marzia, Filice, Marco, Olea, David, Velez, Marisela, Guisan, Jose M and Mateo, Cesar
Abstract:
The preparation and performance of a suitable chimeric biosensor based on antibodies (Abs) immobilized on lipase-coated magnetic particles by means of a standing orienting strategy are presented. This novel system is based on hydrophobic magnetic particles coated with modified lipase molecules able to orient and further immobilize different Abs in a covalent way without any previous site-selective chemical modification of biomacromolecules. Different key parameters attending the process were studied and optimized. The optimal preparation was performed using a controlled loading (1 nmol Ab g(-1) chimeric support) at pH 9 and a short reaction time to recover a biological activity of about 80%. AFM microscopy was used to study and confirm the Abs-oriented immobilization on lipase-coated magnetic particles and the final achievement of a highly active and recyclable chimeric immune sensor. This direct technique was demonstrated to be a powerful alternative to the indirect immunoactivity assay methods for the study of biomacromolecule-oriented immobilizations.
Reference:
Useful oriented immobilization of antibodies on chimeric magnetic particles: direct correlation of biomacromolecule orientation with biological activity by AFM studies. (Marciello, Marzia, Filice, Marco, Olea, David, Velez, Marisela, Guisan, Jose M and Mateo, Cesar), In Langmuir, American Chemical Society, volume 30, 2014.
Bibtex Entry:
@article{Marciello:2014ht,
author = {Marciello, Marzia and Filice, Marco and Olea, David and Velez, Marisela and Guisan, Jose M and Mateo, Cesar},
title = {{Useful oriented immobilization of antibodies on chimeric magnetic particles: direct correlation of biomacromolecule orientation with biological activity by AFM studies.}},
journal = {Langmuir},
year = {2014},
volume = {30},
number = {49},
pages = {15022--15030},
month = dec,
publisher = {American Chemical Society},
affiliation = {Departamento de Biomateriales y Materiales Bioinspirados, Instituto de Ciencia de Materiales de Madrid (ICMM-CSIC), Campus UAM Cantoblanco , 28049 Madrid, Spain.},
doi = {10.1021/la502972v},
pmid = {25420004},
language = {English},
rating = {0},
date-added = {2016-12-12T13:04:21GMT},
date-modified = {2018-03-16T13:14:13GMT},
abstract = {The preparation and performance of a suitable chimeric biosensor based on antibodies (Abs) immobilized on lipase-coated magnetic particles by means of a standing orienting strategy are presented. This novel system is based on hydrophobic magnetic particles coated with modified lipase molecules able to orient and further immobilize different Abs in a covalent way without any previous site-selective chemical modification of biomacromolecules. Different key parameters attending the process were studied and optimized. The optimal preparation was performed using a controlled loading (1 nmol Ab g(-1) chimeric support) at pH 9 and a short reaction time to recover a biological activity of about 80%. AFM microscopy was used to study and confirm the Abs-oriented immobilization on lipase-coated magnetic particles and the final achievement of a highly active and recyclable chimeric immune sensor. This direct technique was demonstrated to be a powerful alternative to the indirect immunoactivity assay methods for the study of biomacromolecule-oriented immobilizations.},
url = {http://pubs.acs.org/doi/abs/10.1021/la502972v},
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uri = {url{papers3://publication/doi/10.1021/la502972v}}
}