Aptamer-Assisted Protein Orientation on Silver Magnetic Nanoparticles: Application to Sensitive Leukocyte Cell-Derived Chemotaxin 2 Surface Plasmon Resonance Sensors

Leukocyte cell-derived chemotaxin 2 (LECT2) has been proved to be a potential biomarker for the diagnosis of liver fibrosis. In this work, a sensitive surface plasmon resonance (SPR) assay for LECT2 analysis was developed. Tyrosine kinase with immune globulin-like and epidermal growth factor-like domains 1 (Tie1) is an orphan receptor of LECT2 with a C-terminal Fc tag, which is far away from the LECT2 binding sites. The Fc aptamer was intentionally used to capture the Tie1 through its Fc tag, connecting with Fe3O4-coated silver magnetic nanoparticles (Ag@MNPs) and ensuring the LECT2 binding site to be outward. Attributed to the orientation nature of the captured protein, Ag@MNPs were able to enhance the SPR signal. A sensitive LECT2 sensor was successfully fabricated with a detection limit of 10.93 pg/mL. The results showed that the immobilization method improved the binding efficiency of Tie1 protein. This strategy could be extended to attach antibodies or recombinant Fc label proteins to Fc aptamer-based nanoparticles.

Publication year: 2022
Authors: Zhu H. 1, Lu Y. 1, Xia J. 1, Liu Y. 2 3, Chen J.1 2, Lee J. 4, Koh K. 5, Chen H. 1 6
Affiliations:
  1. Center for Molecular Recognition and Biosensing, School of Life Sciences, Shanghai University, Shanghai 200444, P.R. China.
  2. School of Medicine, Shanghai University, Shanghai 200444, China.
  3. School of Environmental and Chemical Engineering, Shanghai University, Shanghai 200444, P.R. China.
  4. Department of Chemistry, Chungnam National University, Daejeon 301-747, Republic of Korea.
  5. Institute of General Education, Pusan National University, Busan 609-735, Republic of Korea.
  6. Shanghai Key Laboratory of Bio-Energy Crop, School of Life Sciences, Shanghai University, Shanghai 200444, P.R. China.
Published in: Analytical Chemistry, 2022, Vol. 94, p. 2109-2118
DOI: 10.1021/acs.analchem.1c04448

MP-SPR KEYWORDS

layer-by-layer assembly MP-SPR layer detection protein-polysaccharide interaction

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