title: Biomolecule screening for efficient attachment of biofunctionalized microparticles to the zona pellucida of mammalian oocytes and embryos.
abstract: Individual tagging of oocytes and embryos through the attachment of micrometer-sized polysilicon barcodes to their zona pellucida (ZP) is a promising approach to ensure their correct identification and traceability in human assisted reproduction and in animal production programs. To provide barcodes with the capacity of binding to the ZP, they must be first biofunctionalized with a biomolecule capable of binding to the ZP of both oocytes and embryos. The aim of this work was to select, among an anti-ZP2 antibody and the two lectins wheat germ agglutinin (WGA) and phytohemagglutinin-L, the most optimal biomolecule for the eventual biofunctionalization of barcodes, using mouse oocytes and embryos and commercially available microspheres as a model. Despite the anti-ZP2 antibody showed the highest number of binding sites onto the ZP surface, as determined by field emission scanning electron microscopy, the binding of anti-ZP2-biofunctionalized microspheres to the ZP of cultured oocytes and embryos was less robust and less stable than the binding of lectin-biofunctionalized ones. WGA proved to be, among the three candidates tested, the most appropriate biomolecule to biofunctionalize microparticles with the aim to attach them to the ZP of both oocytes and embryos and to maintain them attached through oocyte activation (zona reaction) and in vitro culture up to the blastocyst stage. As saccharides recognized by WGA are highly abundant in the ZP of most mammalian species, WGA-biofuncionalized microparticles would be able to attach to the ZP of oocytes/embryos of species other than the mouse, such as humans and farm animals.
authors.affiliation: Departament de Biologia Cel lular, Fisiologia i Immunologia, Universitat Autònoma de Barcelona, Facultat Biociències, 08193 Bellaterra, Spain.
authors.forename: Sergio
authors.initials: S
authors.lastname: Novo
authors.forename: Elena
authors.initials: E
authors.lastname: Ibáñez
authors.forename: Leonardo
authors.initials: L
authors.lastname: Barrios
authors.forename: Onofre
authors.initials: O
authors.lastname: Castell
authors.forename: Carme
authors.initials: C
authors.lastname: Nogués
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry; D001665:Binding Sites; D004622:Embryo, Mammalian / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D005260:Female; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008808:Mice, Inbred CBA; D008855:Microscopy, Electron, Scanning; D008863:Microspheres; D009865:Oocytes / Q000187:drug effects / Q000378:metabolism*; D010835:Phytohemagglutinins / Q000737:chemistry / Q000633:toxicity; D013194:Staining and Labeling / Q000379:methods*; D014909:Wheat Germ Agglutinins / Q000737:chemistry / Q000633:toxicity; D015044:Zona Pellucida / Q000378:metabolism*
chemical_list: D000911:Antibodies, Monoclonal; D010835:Phytohemagglutinins; D014909:Wheat Germ Agglutinins