Stable Bioactive Enzyme-Containing Multilayer Films Based on Covalent Cross-Linking from Mussel-Inspired Adhesives.
The use of immobilized enzymes is mandatory for the easy separation of the enzyme, the unreacted substrates, and the obtained products to allow repeated enzymatic assays without cumbersome purification steps. The immobilization procedure is however critical to obtain a high fraction of active enzyme. In this article, we present an enzyme immobilization strategy based on a catechol functionalized alginate. We demonstrate that alkaline phosphatase (ALP) remains active in multilayered films made with alginate modified with catechol moieties (AlgCat) for long duration, that is, up to 7 weeks, provided the multilayered architecture is cross-linked with sodium periodate. This cross-linking reaction allows to create covalent bonds between the amino groups of ALP and the quinone group carried by the modified alginate. In the absence of cross-linking, the enzymatic activity is rapidly lost and this reduction is mainly due to enzyme desorption. We also show that NaIO4 cross-linked (AlgCat-Alp)n films can be freeze-dried and reused at least 3 weeks later without lost in enzymatic activity.
Institut Charles Sadron, Centre National de la Recherche Scientifique, Université de Strasbourg, UPR 22 , 23 rue du Loess, BP 84047, 67034, Strasbourg Cedex 2, France.
Johan
J
Longo
Institut Charles Sadron, Centre National de la Recherche Scientifique, Université de Strasbourg, UPR 22 , 23 rue du Loess, BP 84047, 67034, Strasbourg Cedex 2, France.
Tony
T
Garnier
Biomatériaux et Bioingénierie, INSERM, UMR-S 1121 , 11 rue Humann, 67085 Strasbourg Cedex, France.|Faculté de Chirurgie Dentaire, Université de Strasbourg , 8 rue Sainte Elisabeth. 67000 Strasbourg Cedex, France.
Mihaela
M
Mateescu
Biomatériaux et Bioingénierie, INSERM, UMR-S 1121 , 11 rue Humann, 67085 Strasbourg Cedex, France.|Faculté de Chirurgie Dentaire, Université de Strasbourg , 8 rue Sainte Elisabeth. 67000 Strasbourg Cedex, France.
Florian
F
Ponzio
Institut Charles Sadron, Centre National de la Recherche Scientifique, Université de Strasbourg, UPR 22 , 23 rue du Loess, BP 84047, 67034, Strasbourg Cedex 2, France.|Biomatériaux et Bioingénierie, INSERM, UMR-S 1121 , 11 rue Humann, 67085 Strasbourg Cedex, France.|Faculté de Chirurgie Dentaire, Université de Strasbourg , 8 rue Sainte Elisabeth. 67000 Strasbourg Cedex, France.|Ecole de Chimie, Polymères et Matériaux, Université de Strasbourg , 25 rue Becquerel, 67087 Strasbourg, France.|Institut d'Etudes Avancées de l'Université de Strasbourg , 5 allée du Général Rouvillois, 67083 Strasbourg, France.|International Center for Frontier Research in Chemistry , 8 allée Gaspard Monge, 67083 Strasbourg, France.|Institut Universitaire de France , 103 boulevard Saint-Michel, 75005 Paris, France.
Pierre
P
Schaaf
Institut Charles Sadron, Centre National de la Recherche Scientifique, Université de Strasbourg, UPR 22 , 23 rue du Loess, BP 84047, 67034, Strasbourg Cedex 2, France.|Ecole de Chimie, Polymères et Matériaux, Université de Strasbourg , 25 rue Becquerel, 67087 Strasbourg, France.|Institut d'Etudes Avancées de l'Université de Strasbourg , 5 allée du Général Rouvillois, 67083 Strasbourg, France.|International Center for Frontier Research in Chemistry , 8 allée Gaspard Monge, 67083 Strasbourg, France.
Loïc
L
Jierry
Biomatériaux et Bioingénierie, INSERM, UMR-S 1121 , 11 rue Humann, 67085 Strasbourg Cedex, France.|Faculté de Chirurgie Dentaire, Université de Strasbourg , 8 rue Sainte Elisabeth. 67000 Strasbourg Cedex, France.
Vincent
V
Ball
D000269:Adhesives / Q000737:chemistry*; D000464:Alginates / Q000737:chemistry*; D000469:Alkaline Phosphatase / Q000737:chemistry*; D000818:Animals; D040761:Biomimetic Materials / Q000737:chemistry*; D049872:Bivalvia / Q000737:chemistry / Q000502:physiology; D002396:Catechols / Q000737:chemistry*; D003432:Cross-Linking Reagents / Q000737:chemistry; D057075:Enzyme Assays; D004800:Enzymes, Immobilized / Q000737:chemistry*; D017743:Equipment Reuse; D005612:Freeze Drying; D020723:Glucuronic Acid / Q000737:chemistry; D006603:Hexuronic Acids / Q000737:chemistry; D007700:Kinetics; D010504:Periodic Acid / Q000737:chemistry
D000269:Adhesives; D000464:Alginates; D002396:Catechols; D003432:Cross-Linking Reagents; D004800:Enzymes, Immobilized; D006603:Hexuronic Acids; D010504:Periodic Acid; D020723:Glucuronic Acid; C009288:metaperiodate; D000469:Alkaline Phosphatase