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Directed self-assembly of gold binding polypeptide-protein A fusion proteins for development of gold nanoparticle-based SPR immunosensors.
Biosens Bioelectron. 2009 Apr 15; 24(8):2592-7.BB

Abstract

Effective immobilization of antibodies on a sensing platform and sensitivity enhancement are crucial in designing surface plasmon resonance (SPR) immunosensors. Colloidal gold nanoparticles (AuNPs) were directly assembled onto a surface of SPR Au chip via 2-aminoethanethiol for the enhancement of sensitivity as a label-free detection system. SEM image showed most AuNPs were uniformly distributed over the surface. A novel fusion protein was constructed by genetically fusing gold binding polypeptides (GBP) to protein A (ProA) as a crosslinker for effective immobilization of antibodies. The resulting GBP-ProA protein was directly self-immobilized onto both bare and AuNPs-assembled SPR chip surfaces via the GBP portion, followed by the oriented binding of human immunoglobulin G (hIgG) onto the ProA domain targeting the Fc region of antibodies and anti-hIgG in series. Furthermore, anti-Salmonella antibodies were immobilized onto both GBP-ProA layered chips for detection of Salmonella typhimurium. SPR analyses indicated the signal increases for successive binding of hIgG and anti-hIgG onto the GBP-ProA layered AuNPs-assembled chip were higher (about 92 and 30%, respectively) than that onto the identically treated bare chip. This signal enhancement in the AuNPs-assembled chip also caused a 10-fold increased sensitivity in detection of S. typhimurium compared to the bare one. These results demonstrate the direct assembly of AuNPs onto a SPR chip could enhance the signal in biomolecular interaction events, and the GBP-ProA protein could be a valuable crosslinker for simple and oriented immobilization of antibodies onto Au chip surfaces without any surface chemical modification.

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Publisher Full Text (DOI)

Authors+Show Affiliations

Ko S
Korea Food Research Institute, Seongnam, Gyeonggi-Do 463-746, Republic of Korea. shko7@kfri.re.kr
Park TJ
No affiliation info available
Kim HS
No affiliation info available
Kim JH
No affiliation info available
Cho YJ
No affiliation info available

MeSH

Biosensing TechniquesColony Count, MicrobialCrystallizationEquipment DesignEquipment Failure AnalysisGoldImmunoassayNanoparticlesPeptidesRecombinant Fusion ProteinsReproducibility of ResultsSalmonella typhimuriumSensitivity and SpecificityStaphylococcal Protein ASurface Plasmon Resonance

Pub Type(s)

Evaluation Study
Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

19243930

Citation

Ko, Sungho, et al. "Directed Self-assembly of Gold Binding Polypeptide-protein a Fusion Proteins for Development of Gold Nanoparticle-based SPR Immunosensors." Biosensors & Bioelectronics, vol. 24, no. 8, 2009, pp. 2592-7.
Ko S, Park TJ, Kim HS, et al. Directed self-assembly of gold binding polypeptide-protein A fusion proteins for development of gold nanoparticle-based SPR immunosensors. Biosens Bioelectron. 2009;24(8):2592-7.
Ko, S., Park, T. J., Kim, H. S., Kim, J. H., & Cho, Y. J. (2009). Directed self-assembly of gold binding polypeptide-protein A fusion proteins for development of gold nanoparticle-based SPR immunosensors. Biosensors & Bioelectronics, 24(8), 2592-7. https://doi.org/10.1016/j.bios.2009.01.030
Ko S, et al. Directed Self-assembly of Gold Binding Polypeptide-protein a Fusion Proteins for Development of Gold Nanoparticle-based SPR Immunosensors. Biosens Bioelectron. 2009 Apr 15;24(8):2592-7. PubMed PMID: 19243930.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Directed self-assembly of gold binding polypeptide-protein A fusion proteins for development of gold nanoparticle-based SPR immunosensors. AU - Ko,Sungho, AU - Park,Tae Jung, AU - Kim,Hyo-Sop, AU - Kim,Jae-Ho, AU - Cho,Yong-Jin, Y1 - 2009/01/31/ PY - 2008/10/31/received PY - 2009/01/08/revised PY - 2009/01/09/accepted PY - 2009/2/27/entrez PY - 2009/2/27/pubmed PY - 2009/5/27/medline SP - 2592 EP - 7 JF - Biosensors & bioelectronics JO - Biosens Bioelectron VL - 24 IS - 8 N2 - Effective immobilization of antibodies on a sensing platform and sensitivity enhancement are crucial in designing surface plasmon resonance (SPR) immunosensors. Colloidal gold nanoparticles (AuNPs) were directly assembled onto a surface of SPR Au chip via 2-aminoethanethiol for the enhancement of sensitivity as a label-free detection system. SEM image showed most AuNPs were uniformly distributed over the surface. A novel fusion protein was constructed by genetically fusing gold binding polypeptides (GBP) to protein A (ProA) as a crosslinker for effective immobilization of antibodies. The resulting GBP-ProA protein was directly self-immobilized onto both bare and AuNPs-assembled SPR chip surfaces via the GBP portion, followed by the oriented binding of human immunoglobulin G (hIgG) onto the ProA domain targeting the Fc region of antibodies and anti-hIgG in series. Furthermore, anti-Salmonella antibodies were immobilized onto both GBP-ProA layered chips for detection of Salmonella typhimurium. SPR analyses indicated the signal increases for successive binding of hIgG and anti-hIgG onto the GBP-ProA layered AuNPs-assembled chip were higher (about 92 and 30%, respectively) than that onto the identically treated bare chip. This signal enhancement in the AuNPs-assembled chip also caused a 10-fold increased sensitivity in detection of S. typhimurium compared to the bare one. These results demonstrate the direct assembly of AuNPs onto a SPR chip could enhance the signal in biomolecular interaction events, and the GBP-ProA protein could be a valuable crosslinker for simple and oriented immobilization of antibodies onto Au chip surfaces without any surface chemical modification. SN - 1873-4235 UR - https://www.unboundmedicine.com/medline/citation/19243930/Directed_self_assembly_of_gold_binding_polypeptide_protein_A_fusion_proteins_for_development_of_gold_nanoparticle_based_SPR_immunosensors_ DB - PRIME DP - Unbound Medicine ER -