Journal
MICROCHIMICA ACTA
Volume 183, Issue 1, Pages 1-19Publisher
SPRINGER WIEN
DOI: 10.1007/s00604-015-1623-4
Keywords
Enzyme immobilization; Antibody immobilization; IgG-binding protein; DNA-directed immobilization; Enzyme fusion; Carbohydrate affinity immobilization; Molecular imprinting
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Funding
- National Natural Science Foundation of China [21475106, 31200604]
- Natural Science Foundation of Chongqing [cstc2012jjA10152]
- Fundamental Research Funds for Central Universities [XDJK2015B015]
- Chongqing Key Laboratory for Advanced Materials and Technologies of Clean Energies [cstc2011pt]
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Immobilization of proteins on a solid support is critical with respect to the fabrication and performance of biosensors and biochips. Protein attachment with a preferable orientation can effectively avoid its denaturation and keeps its active sites fully exposed to solution, thus maximally preserving the bioaffinity or bioactivity. This review (with 140 refs.) summarises the recent advances in oriented immobilization of proteins with a particular focus on antibodies and enzymes. Following an introduction that describes reasons for oriented immobilization on (nano)surfaces, we summarize (a) methods for (bio)chemical affinity-mediated oriented immobilization (with sections on immunoglobulin G (IgG)-binding protein as the capture ligand, DNA-directed immobilization, aptamer- and peptide-mediated immobilization, affinity ligand and fusion tag-mediated immobilization, material-binding peptide-assisted immobilization); (b) methods for covalent oriented immobilization (with sections on immobilization via cysteine residues or cysteine tags, via carbohydrate moieties; via enzyme fusion or enzymatic catalysis, and via nucleotide binding sites of antibodies); (c) methods based on molecular imprinting techniques; (d) methods for characterization of oriented immobilized proteins; and then make conclusions and give perspectives.
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