Microchimica Acta

, 186:780 | Cite as

Preconcentration and SERS-based determination of infliximab in blood by using a TNF-α-modified gold-coated copper oxide nanomaterial

  • Saiqa Muneer
  • Godwin A. Ayoko
  • Nazrul Islam
  • Emad L. IzakeEmail author
Original Paper


Infliximab (INF) is a chimeric monoclonal immunoglobulin acting against tumor necrosis factor-alpha (TNF-α). The drug is used for the treatment of chronic autoimmune and inflammatory diseases. A target-specific nanomaterial is presented for the extraction of INF from human plasma along with a label-free surface enhanced Raman spectroscopy (SERS) method for its determination using a handheld device. A gold-coated copper oxide chip was functionalized with TNF-α and used to extract the drug from plasma. INF was recovered from the extractor by lowering the pH value to 2.5. The disulfide bond structure of the drug was then reduced and used for its oriented chemisorption onto a gold-coated copper oxide substrate for SERS measurements using the INF-specific band at 936 cm−1. The working range of the SERS method was between 10−7 and 10−14 M of reduced INF. The relative standard deviation (RSD), between three different measurements was 4.2% (intra-day) and 7.1% (inter-day). The quantification and detection limits of the assay (LOQ, LOD) were 0.01 pM and 1.4 fM respectively. The SERS detection was cross-validated against ELISA where 99% agreement was found between the two methods.

Graphical abstract

Schematic representation of the determination of Infliximab (INF) in blood. A gold coated copper oxide chip was functionalised with tumor necrosis factor (TNF-α) and used to extract INF from blood plasma. The captured INF was released, reduced, chemisorbed onto a second gold-coated copper oxide substrate and screened by surface-enhanced Raman spectroscopy (SERS) using a handheld device.


Functionalised nanomaterial Tumor necrosis factor Disulfide bond structure TNF inhibitor Therapeutic drug monitoring 



We thank FLEW Solutions Australia for donating gold coated copper oxide substrates for this work.

Supplementary material

604_2019_3947_MOESM1_ESM.docx (1.6 mb)
ESM 1 (DOCX 1687 kb)


  1. 1.
    Lichtenstein GR (2013) Comprehensive review: antitumor necrosis factor agents in inflammatory bowel disease and factors implicated in treatment response. Ther Adv Gastroenter 6:269–293CrossRefGoogle Scholar
  2. 2.
    Kirman I, Whelan RL, Nielsen OH (2004) Infliximab: mechanism of action beyond TNF-α neutralization in inflammatory bowel disease. Eur J Gastroen Hepat 16:639–641CrossRefGoogle Scholar
  3. 3.
    Elgundi Z, Reslan M, Cruz E, Sifniotis V, Kayser V (2017) The state-of-play and future of antibody therapeutics. Adv Drug Deliv Rev 122:2–19CrossRefGoogle Scholar
  4. 4.
    Baert F, Noman M, Vermeire S, Van Assche G, D'haens G, Carbonez A, Rutgeerts P (2013) Influence of immunogenicity on the long-term efficacy of infliximab in Crohn's disease. N Engl J Med 348:601–608CrossRefGoogle Scholar
  5. 5.
    Paul S, Moreau AC, Del Tedesco E, Rinaudo M, Phelip J-M, Genin C, Peyrin-Biroulet L, Roblin X (2014) Pharmacokinetics of adalimumab in inflammatory bowel diseases: a systematic review and meta-analysis. Inflamm Bowel Dis 20:1288–1295CrossRefGoogle Scholar
  6. 6.
    Fidder H, Schnitzler F, Ferrante M, Noman M, Katsanos K, Segaert S, Henckaerts L, Van Assche G, Vermeire S, Rutgeerts P (2009) Long-term safety of infliximab for the treatment of inflammatory bowel disease: a single-centre cohort study. Gut 58:501–508CrossRefGoogle Scholar
  7. 7.
    Aarden L, Ruuls SR, Wolbink G (2008) Immunogenicity of anti-tumor necrosis factor antibodies—toward improved methods of anti-antibody measurement. Curr Opin Immunol 20:431–435CrossRefGoogle Scholar
  8. 8.
    Casteele NV, Ferrante M, Van Assche G, Ballet V, Compernolle G, Van Steen K, Simoens S, Rutgeerts P, Gils A, Vermeire S (2015) Trough concentrations of infliximab guide dosing for patients with inflammatory bowel disease. Gastroenterology 148:1320–1329.e1323CrossRefGoogle Scholar
  9. 9.
    Lee MWM, Connor S, Ng W, Toong CM-L (2016) Comparison of infliximab drug measurement across three commercially available ELISA kits. Pathology 38:608–612CrossRefGoogle Scholar
  10. 10.
    Wang S-L, Ohrmund L, Hauenstein S, Salbato J, Reddy R, Monk P, Lockton S, Ling N, Singh S (2012) Development and validation of homogeneous mobility shift assay for measurement of infliximab and antibodies to infliximab levels in patient serum. J Immunol Methods 382:177–188CrossRefGoogle Scholar
  11. 11.
    Lu J, Spasic D, Delport F, Van Stappen T, Deterz I, Daems D, Vermeire S, Gils A, Lammertyn J (2017) Immunoassay for detection of infliximab in whole blood using a fibre-optic surface Plasmon resonance biosensor. Anal Chem 89:3664–3671CrossRefGoogle Scholar
  12. 12.
    Steenholdt C, Ainsworth MA, Tovey M, Klausen TW, Thomsen OØ, Brynskov J, Bendtzen K (2013) Comparison of techniques for monitoring infliximab and antibodies against infliximab in crohn's disease. Ther Drug Monit 35:530–538CrossRefGoogle Scholar
  13. 13.
    Sharma B, Frontiera RR, Henry A-I, Ringe E, Van Duyne RP (2012) SERS: materials, applications, and the future. Mater Today 15:16–25CrossRefGoogle Scholar
  14. 14.
    Zhang W, Jiang L, Piper JA, Wang Y (2018) SERS nanotags and their applications in biosensing and bioimaging. J Anal Test 2:26–44CrossRefGoogle Scholar
  15. 15.
    Freeman LM, Pang L, Fainman Y (2014) Maximizing the electromagnetic and chemical resonances of surface-enhanced Raman scattering for nucleic acids. ACS Nano 8:8383–8391CrossRefGoogle Scholar
  16. 16.
    Ding S-Y, You E-M, Tian Z-Q, Moskovits M (2017) Electromagnetic theories of surface-enhanced Raman spectroscopy. Chem Soc Rev 46(13):4042–4076CrossRefGoogle Scholar
  17. 17.
    Zheng X-S, Jahn IJ, Weber K, Cialla-May D, Popp J (2018) Label-free SERS in biological and biomedical applications: recent progress, current challenges and opportunities. Spectrochim Acta A Mol Biomol Spectrosc 197:56–77CrossRefGoogle Scholar
  18. 18.
    Pilot R, Signorini R, Durante C, Orian L, Bhamidipati M, Fabris L (2019) A review on surface-enhanced Raman scattering. Biosensors 9(57). CrossRefGoogle Scholar
  19. 19.
    Lastek (2019) Accessed 14 Oct 2019
  20. 20.
    Hassanain WA, Izake EL, Schmidt MS, Ayoko GA (2017) Gold nanomaterials for the selective capturing and SERS diagnosis of toxins in aqueous and biological fluids. Biosens Bioelectron 91:664–672CrossRefGoogle Scholar
  21. 21.
    Agoston R, Izake EL, Sivanesan A, Lott WB, Sillence M, Steel R (2016) Rapid isolation and detection of erythropoietin in blood plasma by magnetic core gold nanoparticles and portable Raman spectroscopy. Nanomed Nanotechnol Biol Med 12:633–641CrossRefGoogle Scholar
  22. 22.
    Balčytis A, Ryu M, Seniutinas G, Juodkazytė J, Cowie BC, Stoddart PR, Zamengo M, Morikawa J, Juodkazis S (2015) Black-CuO: surface-enhanced Raman scattering and infrared properties. Nanoscale 7:18299–304.sCrossRefGoogle Scholar
  23. 23.
  24. 24.
    Xu W, Ling X, Xiao J, Dresselhaus MS, Kong J, Xu H, Liu Z, Zhang J (2012) Surface enhanced Raman spectroscopy on a flat graphene surface. Proc Natl Acad Sci 109:9281–9286CrossRefGoogle Scholar
  25. 25.
    Hassanain WA, Izake EL, Ayoko GA (2018) Spectroelectrochemical nanosensor for the determination of cystatin C in human blood. Anal Chem 90:10843–10850CrossRefGoogle Scholar
  26. 26.
    Park W-H, Kim ZH (2010) Charge transfer enhancement in the SERS of a single molecule. Nano Lett 10:4040–4048CrossRefGoogle Scholar
  27. 27.
    Demirel G, Usta H, Yilmaz M, Celik M, Alidagi HA, Buyukserin F (2018) Surface-enhanced Raman spectroscopy (SERS): an adventure from plasmonic metals to organic semiconductors as SERS platforms. J Mater Chem C 6:5314–5335CrossRefGoogle Scholar
  28. 28.
    Boyaci IH, Temiz HT, Geniş HE, Soykut EA, Yazgan NN, Güven B, Uysal RS, Bozkurt AG, İlaslan K, Toruna O, Şeker FCD (2015) Dispersive and FT-Raman spectroscopic methods in food analysis. RSC Adv 5:56606–56624CrossRefGoogle Scholar
  29. 29.
    Rygula A, Majzner K, Marzec KM, Kaczor A, Pilarczyk M, Baranska M (2013) Raman spectroscopy of proteins: a review. J Raman Spectrosc 44:1061–1076CrossRefGoogle Scholar
  30. 30.
    Sanles-Sobrido M, Rodríguez-Lorenzo L, Lorenzo-Abalde S, González-Fernández Á, Correa-Duarte MA, Alvarez-Puebla RA, Liz-Marzán LM (2009) Label-free SERS detection of relevant bioanalytes on silver-coated carbon nanotubes: the case of cocaine. Nanoscale 1:153–158CrossRefGoogle Scholar
  31. 31.
    Podstawka E, Ozaki Y, Proniewicz LM (2004) Part I: surface-enhanced Raman spectroscopy investigation of amino acids and their homodipeptides adsorbed on colloidal silver. Appl Spectrosc 58:570–580CrossRefGoogle Scholar
  32. 32.
    Sivanesan A, Izake EL, Agoston R, Ayoko GA, Sillence M (2015) Reproducible and label-free biosensor for the selective extraction and rapid detection of proteins in biological fluids. J Nanobiotechnol 13:43CrossRefGoogle Scholar
  33. 33.
    Mauser JF, Hyams JS (1999) Infliximab: a novel chimeric monoclonal antibody for the treatment of Crohn's disease. Clin Ther 21:932–942CrossRefGoogle Scholar
  34. 34.
    Chapman RG, Ostuni E, Yan L, Whitesides GM (2000) Preparation of mixed self-assembled monolayers (SAMs) that resist adsorption of proteins using the reaction of amines with a SAM that presents interchain carboxylic anhydride groups. Langmuir 16:6927–6936CrossRefGoogle Scholar
  35. 35.
    Kausaite-Minkstimiene A, Ramanaviciene A, Kirlyte J, Ramanavicius A (2010) Comparative study of random and oriented antibody immobilization techniques on the binding capacity of immunosensor. Anal Chem 82:6401–6408CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Austria, part of Springer Nature 2019

Authors and Affiliations

  • Saiqa Muneer
    • 1
    • 2
  • Godwin A. Ayoko
    • 1
    • 2
  • Nazrul Islam
    • 2
    • 3
  • Emad L. Izake
    • 1
    • 2
    Email author
  1. 1.Molecular Design and Synthesis Discipline, Science and Engineering FacultyQueensland University of TechnologyBrisbaneAustralia
  2. 2.Discipline of Environmental Technologies, Science and Engineering FacultyQueensland University of TechnologyBrisbaneAustralia
  3. 3.School of Clinical Sciences, Faculty of HealthQueensland University of TechnologyBrisbaneAustralia

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