Microchimica Acta

, 185:240 | Cite as

The mechanism of the adsorption of dsDNA on citrate-stabilized gold nanoparticles and a colorimetric and visual method for detecting the V600E point mutation of the BRAF gene

  • Zhen Liu
  • Menik Hettihewa
  • Yang Shu
  • Chunqiong Zhou
  • Qian Wan
  • Lihong Liu
Original Paper

Abstract

A study is presented on the binding kinetics and mechanism of the adsorption of dsDNA on citrate-capped gold nanoparticles (AuNPs). Methods include fluorescence titration, isothermal calorimetry (ITC) titration, dynamic light scattering and gel electrophoresis. It is found that the fluorescence of probe DNA (labeled with Rhodamine Green and measured at excitation/emission peaks of 498/531 nm) is quenched by addition of AuNPs. The Stern-Volmer quenching constant (Ksv) is 1.67 × 10^9 L·mol−1 at 308 K and drops with increasing temperature. The quenching mechanism is mainly static. The results of both fluorescence titrations and ITC show negative values for ΔH and ΔS values. This shows ion-induced dipole-dipole interaction to be the main attractive forces between dsDNA and AuNPs, while electrostatic interactions result in repulsion. The repulsive forces lead to a lower affinity between dsDNA and AuNPs (compared to single-strand DNA). It is also found that dsDNA can prevent the aggregation of AuNPs which is accompanied by a color change from red into blue. The visual detection limit with bare eyes for dsDNA1 is 36 pM. Based on these findings, a colorimetric method was developed to detect the proto-oncogene of serine/threonine-protein kinase B-Raf V600E point mutation in HT29, Ec109, A549, Huh-7 and SW480 cell lines.

Graphical abstract

Schematic of the salt-induced aggregation of uncapped gold nanoparticles (AuNPs) which leads to a color change from red to blue. If the AuNPs are coated with dsDNA, aggregation is suppressed.

Keywords

Unmodified gold nanoparticles Electrostatic interactions Ion-induced dipole interaction dsDNA Colorimetric detection Mutation detection 

Notes

Acknowledgments

The study was supported financially by the Natural Science Foundation of China (81470161) and the Science and Technology Program of Guangdong Province (2016A040403052 for Liu, 2014A050503042 for Zhou).

Author contributions

L. L and Z. L designed research; Z. L, Y. S, L. L and Q. W performed research, analyzed data; L. L, M. H, Z. L, Y. S and C. Z wrote the paper.

Compliance with ethical standards

The author(s) declare that they have no competing interests.

Supplementary material

604_2018_2775_MOESM1_ESM.docx (6.5 mb)
ESM 1 (DOCX 6612 kb)

References

  1. 1.
    Holzinger M, Le Goff A, Cosnier S (2014) Nanomaterials for biosensing applications: a review. Front Chem 2:63CrossRefGoogle Scholar
  2. 2.
    Elghanian R, Storhoff JJ, Mucic RC, Letsinger RL, Mirkin CA (1997) Selective colorimetric detection of polynucleotides based on the distance-dependent optical properties of gold nanoparticles. Science 277:1078–1081CrossRefGoogle Scholar
  3. 3.
    Mirkin CA, Letsinger RL, Mucic RC, Storhoff JJ (1996) A DNA-based method for rationally assembling nanoparticles into macroscopic materials. Nature 382:607–609CrossRefGoogle Scholar
  4. 4.
    Li H, Rothberg L (2004) Colorimetric detection of DNA sequences based on electrostatic interactions with unmodified gold nanoparticles. Proc Natl Acad Sci U S A 101:14036–14039CrossRefGoogle Scholar
  5. 5.
    Lee JH, Wang Z, Liu J, Lu Y (2008) Highly sensitive and selective colorimetric sensors for uranyl (UO2(2+)): development and comparison of labeled and label-free DNAzyme-gold nanoparticle systems. J Am Chem Soc 130:14217–14226CrossRefGoogle Scholar
  6. 6.
    Seow N, Lai PS, Yung LY (2014) Gold nanostructures for the multiplex detection of glucose-6-phosphate dehydrogenase gene mutations. Anal Biochem 451:56–62CrossRefGoogle Scholar
  7. 7.
    Veigas B, Pedrosa P, Couto I, Viveiros M, Baptista PV (2013) Isothermal DNA amplification coupled to au-nanoprobes for detection of mutations associated to rifampicin resistance in Mycobacterium tuberculosis. J Nanobiotechnology 11:38CrossRefGoogle Scholar
  8. 8.
    Valentini P, Fiammengo R, Sabella S, Gariboldi M, Maiorano G, Cingolani R, Pompa PP (2013) Gold-nanoparticle-based colorimetric discrimination of cancer-related point mutations with picomolar sensitivity. ACS Nano 7:5530–5538CrossRefGoogle Scholar
  9. 9.
    Qin WJ, Yim OS, Lai PS, Yung LY (2010) Dimeric gold nanoparticle assembly for detection and discrimination of single nucleotide mutation in Duchenne muscular dystrophy. Biosens Bioelectron 25:2021–2025CrossRefGoogle Scholar
  10. 10.
    Nelson EM, Rothberg LJ (2011) Kinetics and mechanism of single-stranded DNA adsorption onto citrate-stabilized gold nanoparticles in colloidal solution. Langmuir 27:1770–1777CrossRefGoogle Scholar
  11. 11.
    Sandström P, Mila Boncheva A, Åkerman B (2003) Nonspecific and thiol-specific binding of DNA to gold nanoparticles. Langmuir 19:7537–7543CrossRefGoogle Scholar
  12. 12.
    Cárdenas M, Barauskas J, Schillén K, Brennan JL, Brust M, Nylander T (2006) Thiol-specific and nonspecific interactions between DNA and gold nanoparticles. Langmuir 22:3294–3299CrossRefGoogle Scholar
  13. 13.
    Lin YZ, Chang PL (2014) Colorimetric determination of DNA methylation based on the strength of the hydrophobic interactions between DNA and gold nanoparticles. ACS Appl Mater Inter 5:12045–12051CrossRefGoogle Scholar
  14. 14.
    Farkhari N, Abbasian S, Moshaii A, Nikkhah M (2016) Mechanism of adsorption of single and double stranded DNA on gold and silver nanoparticles: investigating some important parameters in bio-sensing applications. Colloids surf B Biointerfaces 148:657–664CrossRefGoogle Scholar
  15. 15.
    Carnerero JM, Sánchez-Coronilla A, Martín EI, Jimenez-Ruiz A, Prado-Gotor R (2017) Quantification of nucleobases/gold nanoparticles interactions: energetics of the interactions through apparent binding constants determination. Phys Chem Chem Phys 19:16113–16123CrossRefGoogle Scholar
  16. 16.
    Koo KM, Sina AAI, Carrascosa LG, Shiddiky MJA, Trau M (2015) DNA-bare gold affinity interactions: mechanism and applications in biosensing. Anal Methods 7(17):7042–7054CrossRefGoogle Scholar
  17. 17.
    Kovach JS, Hartmann A, Blaszyk H, Cunningham J, Schaid D, Sommer SS (1996) Mutation detection by highly sensitive methods indicates that p53 gene mutations in breast cancer can have important prognostic value. Proc Natl Acad Sci U S A 93:1093–1096CrossRefGoogle Scholar
  18. 18.
    Davies H, Bignell GR, Cox C, Stephens P, Edkins S, Clegg S, Teague J, Woffendin H, Garnett MJ, Bottomley W, Davis N, Dicks E, Ewing R, Floyd Y, Gray K, Hall S, Hawes R, Hughes J, Kosmidou V, Menzies A, Mould C, Parker A, Stevens C, Watt S, Hooper S, Wilson R, Jayatilake H, Gusterson BA, Cooper C, Shipley J, Hargrave D, Pritchard-Jones K, Maitland N, Chenevix-Trench G, Riggins GJ, Bigner DD, Palmieri G, Cossu A, Flanagan A, Nicholson A, Ho JW, Leung SY, Yuen ST, Weber BL, Seigler HF, Darrow TL, Paterson H, Marais R, Marshall CJ, Wooster R, Stratton MR, Futreal PA (2002) Mutations of the BRAF gene in human cancer. Nature 417:949–954CrossRefGoogle Scholar
  19. 19.
    Tang HC, Chen YC (2015) Molecular insight and resolution for tumors harboring the H-ras (G12V) mutation. RSC Adv 5:20623–20633CrossRefGoogle Scholar
  20. 20.
    Strohmeier O, Laßmann S, Riedel B, Mark D, Roth G, Werner M, Zengerle R, Von Stetten F (2014) Multiplex genotyping of KRAS point mutations in tumor cell DNA by allele-specific real-time PCR on a centrifugal microfluidic disk segment. Microchim Acta 181:1681–1688CrossRefGoogle Scholar
  21. 21.
    Lee PC, Meisel D (1982) Adsorption and surface-enhanced Raman of dyes on silver and gold sols. J Phys Chem B 86:3391–3395CrossRefGoogle Scholar
  22. 22.
    Situ B, Cao N, Li B, Liu Q, Lin L, Dai Z, Zou X, Cai Z, Wang Q, Yan X, Zheng L (2013) Sensitive electrochemical analysis of BRAF V600E mutation based on an amplification-refractory mutation system coupled with multienzyme functionalized Fe3O4/au nanoparticles. Biosens Bioelectron 43:257–263CrossRefGoogle Scholar
  23. 23.
    Haiss W, Thanh NT, Aveyard J, Fernig D (2007) Determination of size and concentration of gold nanoparticles from UV-vis spectra. Anal Chem 79:4215–4221CrossRefGoogle Scholar
  24. 24.
    Jarry A, Masson D, Cassagnau E, Parois S, Laboisse C, Denis MG (2004) Real-time allele-specific amplification for sensitive detection of the BRAF mutation V600E. Mol Cell robes 18:349–352CrossRefGoogle Scholar
  25. 25.
    Deng H, Zhang X, Kumar A, Zou G, Zhang X, Liang XJ (2013) Long genomic DNA amplicons adsorption onto unmodified gold nanoparticles for colorimetric detection of bacillus anthracis. Chem Commun (Camb) 49:51–53CrossRefGoogle Scholar
  26. 26.
    Islam MM, Chakraborty M, Pandya P, Masum AA, Gupta N, Mukhopadhyay S (2013) Binding of DNA with rhodamine B: spectroscopic and molecular modeling studies. Dyes Pigments 99:412–422CrossRefGoogle Scholar
  27. 27.
    Pylaev TE, Volkova EK, Kochubey VI, Bogatyrev VA, Khlebtsov NG (2013) DNA detection assay based on fluorescence quenching of rhodamine B by gold nanoparticles: the optical mechanisms. J Quant Spectrosc Ra 131:34–42CrossRefGoogle Scholar
  28. 28.
    Jayabharathi J, Jayamoorthy K, Thanikachalam V (2012) Docking investigation and binding interaction of benzimidazole derivative with bovine serum albumin. J Photochem Photobiol B 117:27–32CrossRefGoogle Scholar
  29. 29.
    Shu Y, Xue W, Xu X, Jia Z, Yao X, Liu S, Liu L (2015) Interaction of erucic acid with bovine serum albumin using a multi-spectroscopic method and molecular docking technique. Food Chem 173:31–37CrossRefGoogle Scholar
  30. 30.
    Zhang J, Zhuang S, Tong C, Liu W (2013) Probing the molecular interaction of triazole fungicides with human serum albumin by multispectroscopic techniques and molecular modeling. J Agric Food Chem 61:7203–7211CrossRefGoogle Scholar
  31. 31.
    Ross PD, Subramanian S (1981) Thermodynamics of protein association reactions: forces contributing to stability. Biochemist 20:3096–3102CrossRefGoogle Scholar
  32. 32.
    Callies O, Hernández Daranas A (2016) Application of isothermal titration calorimetry as a tool to study natural product interactions. Nat Prod Rep 33:881–904CrossRefGoogle Scholar
  33. 33.
    Tol J, Dijkstra JR, Klomp M, Teerenstra S, Dommerholt M, Vink-Börger ME, van Cleef PH, van Krieken JH, Punt CJ, Nagtegaal ID (2010) Markers for EGFR pathway activation as predictor of outcome in metastatic colorectal cancer patients treated with or without cetuximab. Eur J Cancer 46:1997–2009CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Austria, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Guangdong Provincial Key Laboratory of New Drug Screening, School of Pharmaceutical SciencesSouthern Medical UniversityGuangzhouChina
  2. 2.Department of Pharmacology, Faculty of MedicineUniversity of RuhunaGalleSri Lanka

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