Optimized immobilization of lectins using self-assembled monolayers on polysilicon encoded materials for cell tagging.
Self-assembled monolayers (SAMs) have been used for the preparation of functional microtools consisting of encoded polysilicon barcodes biofunctionalized with proteins of the lectin family. These hybrid microtools exploit the lectins ability for recognizing specific carbohydrates of the cell membrane to give an efficient system for cell tagging. This work describes how the control of the methodology for SAM formation on polysilicon surfaces followed by lectin immobilization has a crucial influence on the microtool biofunction. Several parameters (silanization time, silane molar concentration, type of solvent or deposition methodology) have been studied to establish optimal function. Furthermore, silanes incorporating different terminal groups, such as aldehyde, activated ester or epoxide groups were tested in order to analyze their chemical coupling with the biomolecules, as well as their influence on the biofunctionality of the immobilized protein. Two different lectins - wheat germ agglutinin (WGA) and phytohemagglutinin (PHA-L) - were immobilized, because they have different and specific cell recognition behaviour and exhibit different cell toxicity. In this way we can assess the effect of intrinsic bulk toxicity with that of the cell compatibility once immobilized as well as the importance of cell affinity. A variety of nanometrical techniques were used to characterize the active surfaces, and lectin immobilization was quantified using ultraviolet-visible absorption spectroscopy (UV-vis) and optical waveguide light mode spectroscopy (OWLS). Once the best protocol was found, WGA and PHA were immobilized on polysilicon coded barcodes, and these microtools showed excellent cell tagging on living mouse embryos when WGA was used.
Department of Pharmacology and Therapeutical Chemistry and Institute of Nanoscience and Nanotechnology UB (IN2UB), Universitat de Barcelona, Avda. Joan XXIII s/n, 08028 Barcelona, Spain.
Oriol
O
Penon
Department of Pharmacology and Therapeutical Chemistry and Institute of Nanoscience and Nanotechnology UB (IN2UB), Universitat de Barcelona, Avda. Joan XXIII s/n, 08028 Barcelona, Spain.
Dimitrios
D
Siapkas
Department of Cellular Biology, Physiology and Immunology, Universitat Autònoma de Barcelona, 08193 Bellaterra, Spain.
Sergi
S
Novo
Institute of Microelectronics of Barcelona, IMB-CNM (CSIC), Campus UAB, 08193 Bellaterra, Barcelona, Spain.
Sara
S
Durán
Scientific and Technological Centers of the Universitat de Barcelona, Lluís Solé i Sabarís 1, 08028, Barcelona, Spain.
Gerard
G
Oncins
Universite de Lyon, Lyon1, Institut des Sciences Analytiques (ISA), UMR 5280, 5 Rue de la Doua, 69100 Villeurbanne Cedex, France.
Abdelhamid
A
Errachid
Department of Cellular Biology, Physiology and Immunology, Universitat Autònoma de Barcelona, 08193 Bellaterra, Spain.
Lleonard
L
Barrios
Department of Cellular Biology, Physiology and Immunology, Universitat Autònoma de Barcelona, 08193 Bellaterra, Spain.
Carme
C
Nogués
Institute of Microelectronics of Barcelona, IMB-CNM (CSIC), Campus UAB, 08193 Bellaterra, Barcelona, Spain.
Marta
M
Duch
Institute of Microelectronics of Barcelona, IMB-CNM (CSIC), Campus UAB, 08193 Bellaterra, Barcelona, Spain.
José Antonio
JA
Plaza
Department of Pharmacology and Therapeutical Chemistry and Institute of Nanoscience and Nanotechnology UB (IN2UB), Universitat de Barcelona, Avda. Joan XXIII s/n, 08028 Barcelona, Spain. Electronic address: mlperez@ub.edu.
Lluïsa
L
Pérez-García
D000818:Animals; D002448:Cell Adhesion; D002462:Cell Membrane / Q000737:chemistry; D037102:Lectins / Q000737:chemistry*; D051379:Mice; D011108:Polymers / Q000138:chemical synthesis / Q000737:chemistry*; D012825:Silicon / Q000737:chemistry*; D013499:Surface Properties
D037102:Lectins; D011108:Polymers; D012825:Silicon
Cell tagging; Immobilization; Lectins; Polysilicon; Self-assembled monolayers; Surface chemistry