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, volume 30, 2014.
Bibtex Entry:
@article{marciello_useful_2014,
	title = {Useful oriented immobilization of antibodies on chimeric magnetic particles: direct correlation of biomacromolecule orientation with biological activity by {AFM} studies.},
	volume = {30},
	url = {http://pubs.acs.org/doi/10.1021/la502972v},
	doi = {10.1021/la502972v},
	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.},
	language = {English},
	number = {49},
	journal = {Langmuir},
	author = {Marciello, Marzia and Filice, Marco and Olea, David and Velez, Marisela and Guisan, Jose M and Mateo, Cesar},
	month = dec,
	year = {2014},
	pmid = {25420004},
	note = {Publisher: American Chemical Society},
	pages = {15022--15030}
}