Protein Engineering For Directed Immobilization

Abstract

Much effort has been put into the optimization of the functional activity of proteins. For biosensors this protein functional optimization will increase the biosensor's sensitivity and/or selectivity. However, the strategy chosen for the immobilization of the proteins to the sensor surface might be equally important for the development of sensor surfaces that are optimally biologically active. Several studies published in recent years show that the oriented immobilization of the bioactive molecules improves the sensor's properties. In this review, we discuss the state of the art of the different protein immobilization strategies that are commonly used today with a special focus on biosensor applications. These strategies include nonspecific immobilization techniques either by physical adsorption, by covalent coupling, or by specific immobilization via site-specifically introduced tags or bio-orthogonal chemistry. The different tags and bio-orthogonal chemistry available and the techniques to site-specifically introduce these groups in proteins are also discussed.

Bio-orthogonal Immobilization of Fibroblast Growth Factor 2 for Spatial Controlled Cell Proliferation

“…These and other concerns challenge the use of unspecific immobilization chemistries, including EDC/NHS coupling. 9,10 Site-specific chemistries remove these challenges through homogeneous decoration by strictly confining the covalent modification to, for example, an unnatural functional group or groups that are introduced into the biologic's backbone at the predefined site during recombinant protein expression. 11,12 This approach takes advantage of the translation machinery of archaebacteria, which incorporate the 22nd amino acid Lpyrrolysine (Pyl) in protein biosynthesis beyond the typically used 20 natural amino acid building blocks.…”

Development of Cellulose Nanofibre (Cnf) Derived From Kenaf Bast Fibre and It’s Potential in Enzyme Immobilization Support

“…Chemical coupling agents also permit a very strong binding between target enzyme molecules and CNF support at a specific point of reactive group, as promising the accessibility and flexibility of the interaction [18]. Reactive functional group acts as a platform for the interaction between two end-terminals of the chemical coupling, providing the highest chance to create bio-functionalized linkage with the amino group of enzyme [19].…”

Centrifugal Microfluidic Control Mechanisms for Biosensors

“…Thus, the improvement of hydrophilicity or hydrophobicity can increase the biosensor's sensitivity and selectivity. (34,35) However, the downside is that the chemically treated agents can dissolve in the sample solution and influence the results of the chemical reaction. …”