title: Useful oriented immobilization of antibodies on chimeric magnetic particles: direct correlation of biomacromolecule orientation with biological activity by AFM studies.
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.
authors.affiliation: Departamento de Biomateriales y Materiales Bioinspirados, Instituto de Ciencia de Materiales de Madrid (ICMM-CSIC), Campus UAM Cantoblanco , 28049 Madrid, Spain.
authors.forename: Marzia
authors.initials: M
authors.lastname: Marciello
authors.forename: Marco
authors.initials: M
authors.lastname: Filice
authors.forename: David
authors.initials: D
authors.lastname: Olea
authors.forename: Marisela
authors.initials: M
authors.lastname: Velez
authors.forename: José M
authors.initials: JM
authors.lastname: Guisan
authors.forename: Cesar
authors.initials: C
authors.lastname: Mateo
mesh_terms: D055910:Antibodies, Immobilized / Q000737:chemistry*; D015374:Biosensing Techniques; D008280:Magnetics; D018625:Microscopy, Atomic Force; D008954:Models, Biological; D008958:Models, Molecular; D010316:Particle Size
chemical_list: D055910:Antibodies, Immobilized