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Oriented immobilization of proteins on solid supports for use in biosensors and biochips: a review

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Abstract

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.

This review summarises recent advances in oriented immobilization of proteins based on strategies via bio−/chemical affinity, covalent bonding, and molecular imprinting techniques. Advantages and disadvantages of each approach are discussed.

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Acknowledgments

This work is financially supported by the National Natural Science Foundation of China (Grant 21475106, 31200604), Natural Science Foundation of Chongqing (Grant cstc2012jjA10152), the Fundamental Research Funds for the Central Universities (XDJK2015B015), Chongqing Key Laboratory for Advanced Materials and Technologies of Clean Energies (Grant cstc2011pt).

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  1. Institute for Clean Energy & Advanced Materials, Faculty of Materials & Energy, Southwest University, Chongqing, 400715, China

    Yingshuai Liu & Jie Yu

  2. Chongqing Engineering Research Center for Rapid Diagnosis of Dread Disease, Southwest University, Chongqing, 400715, China

    Yingshuai Liu & Jie Yu

  3. Chongqing Key Laboratory for Advanced Materials and Technologies of Clean Energies, Southwest University, Chongqing, 400715, China

    Yingshuai Liu & Jie Yu

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  1. Yingshuai Liu
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  2. Jie Yu
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Correspondence to Yingshuai Liu.

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Liu, Y., Yu, J. Oriented immobilization of proteins on solid supports for use in biosensors and biochips: a review. Microchim Acta 183, 1–19 (2016). https://doi.org/10.1007/s00604-015-1623-4

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