Abstract
An electrochemical approach is presented based on multiwall carbon nanotubes (MWCNTs) and neodymium(III) hydroxide (Nd(OH)3) nanoflakes for detection of bovine serum albumin (BSA). The materials were characterized morphologically (XRPD, SEM, and HR-TEM) and electrochemically (DPV, EIS). The MWCNTs@Nd(OH)3 composite was used as support for bovine serum albumin polyclonal antibody (anti-BSA). After the antibody immobilization on the electrochemical platform and antigen/antibody binding time (optimum 60 min), the proposed approach shows a linear voltammetric response toward BSA concentration in the range 0.066 to 6.010 ng mL−1 at maximum peak potential of 0.13 V (vs. Ag/AgCl). Limit of detection (LOD) and limit of quantification (LOQ) were 18 pg mL−1 and 61 pg mL−1, respectively. The precision of the method calculated as relative standard deviation (RSD) of five independent measurements was better 3%. The selectivity of the optimized method regarding structurally similar proteins (human serum albumin and human hemoglobin), ions (Na+, K+, Ca2+, and NO2−), or compounds (glucose, ascorbic acid, dopamine, uric acid, paracetamol, and glycine) was found to be satisfactory, with the current changes of less than 5% in the presence of up to 1 × 105 times higher concentrations (depending on the compound) of the listed potential interfering compounds. Practical applicability of immunosensor for BSA determination in cow whey sample, with recovery values in the range 97 to 103%, shows that the developed method has high potential for precise and accurate detection of BSA, as well as exceptional miniaturization possibilities for on-site and equipment-free sensing.
Graphical abstract

This is a preview of subscription content, access via your institution.





References
Li Y, Wang X, Wang Y (2006) Comparative studies on interactions of bovine serum albumin with cationic gemini and single-chain surfactants. J Phys Chem B 110:8499–8505. https://doi.org/10.1021/jp060532n
Jahanban-Esfahlan A, Ostadrahimi A, Jahanban-Esfahlan R, Roufegarinejad L, Tabibiazar M, Amarowicz R (2019) Recent developments in the detection of bovine serum albumin. Int J Biol Macromol 138:602–617. https://doi.org/10.1016/j.ijbiomac.2019.07.096
Taghipour P, Zakariazadeh M, Sharifi M, Ezzati Nazhad Dolatabadi J, Barzegar A (2018) Bovine serum albumin binding study to erlotinib using surface plasmon resonance and molecular docking methods. Journal of photochemistry and photobiology. B, Biology 183:11–15. https://doi.org/10.1016/j.jphotobiol.2018.04.008
Tacoma R, Fields J, Ebenstein DB, Lam Y-W, Greenwood SL (2016) Characterization of the bovine milk proteome in early-lactation Holstein and Jersey breeds of dairy cows. J Proteomics 130:200–210. https://doi.org/10.1016/j.jprot.2015.09.024
Restani P, Ballabio C, Di Lorenzo C, Tripodi S, Fiocchi A (2009) Molecular aspects of milk allergens and their role in clinical events. Anal Bioanal Chem 395:47–56. https://doi.org/10.1007/s00216-009-2909-3
Monaci L, Tregoat V, van Hengel AJ, Anklam E (2006) Milk allergens, their characteristics and their detection in food: a review. Eur Food Res Technol 223:149–179. https://doi.org/10.1007/s00217-005-0178-8
Medetalibeyoglu H, Beytur M, Akyıldırım O, Atar N, Yola ML (2020) Validated electrochemical immunosensor for ultra-sensitive procalcitonin detection: carbon electrode modified with gold nanoparticles functionalized sulfur doped MXene as sensor platform and carboxylated graphitic carbon nitride as signal amplification. Sens Actuators, B Chem 319:128195. https://doi.org/10.1016/j.snb.2020.128195
Stanković V, Đurđić S, Ognjanović M, Antić B, Kalcher K, Mutić J, Stanković DM (2020) Anti-human albumin monoclonal antibody immobilized on EDC-NHS functionalized carboxylic graphene/AuNPs composite as promising electrochemical HSA immunosensor. J Electroanal Chem 860:113928. https://doi.org/10.1016/j.jelechem.2020.113928
Garkani Nejad F, Asadi MH, Sheikhshoaie I, Dourandish Z, Zaimbashi R, Beitollahi H (2022) Construction of modified screen-printed graphite electrode for the application in electrochemical detection of sunset yellow in food samples. Food and chemical toxicology an international journal published for the British Industrial Biological Research Association 166:113243. https://doi.org/10.1016/j.fct.2022.113243
Beitollahi H, Shahsavari M, Sheikhshoaie I, Tajik S, Jahani PM, Mohammadi SZ, Afshar AA (2022) Amplified electrochemical sensor employing screen-printed electrode modified with Ni-ZIF-67 nanocomposite for high sensitive analysis of Sudan I in present bisphenol A. Food and chemical toxicology an international journal published for the British Industrial Biological Research Association 161:112824. https://doi.org/10.1016/j.fct.2022.112824
Orooji Y, Asrami PN, Beitollahi H, Tajik S, Alizadeh M, Salmanpour S, Baghayeri M, Rouhi J, Sanati AL, Karimi F (2021) An electrochemical strategy for toxic ractopamine sensing in pork samples; twofold amplified nano-based structure analytical tool. Food Measure 15:4098–4104. https://doi.org/10.1007/s11694-021-00982-y
Tajik S, Beitollahi H, Torkzadeh-Mahani M (2022) Electrochemical immunosensor for the detection of anti-thyroid peroxidase antibody by gold nanoparticles and ionic liquid-modified carbon paste electrode. J Nanostruct Chem 12:581–588. https://doi.org/10.1007/s40097-022-00496-z
Tajik S, Orooji Y, Karimi F, Ghazanfari Z, Beitollahi H, Shokouhimehr M, Varma RS, Jang HW (2021) High performance of screen-printed graphite electrode modified with Ni–Mo-MOF for voltammetric determination of amaranth. Food Measure 15:4617–4622. https://doi.org/10.1007/s11694-021-01027-0
Hartati YW, Letelay LK, Gaffar S, Wyantuti S, Bahti HH (2020) Cerium oxide-monoclonal antibody bioconjugate for electrochemical immunosensing of HER2 as a breast cancer biomarker. Sensing and Bio-Sensing Research 27:100316. https://doi.org/10.1016/j.sbsr.2019.100316
Kadadou D, Tizani L, Wadi VS, Banat F, Naddeo V, Alsafar H, Yousef AF, Hasan SW (2022) Optimization of an rGO-based biosensor for the sensitive detection of bovine serum albumin: effect of electric field on detection capability. Chemosphere 301:134700. https://doi.org/10.1016/j.chemosphere.2022.134700
Kukkar M, Sharma A, Kumar P, Kim K-H, Deep A (2016) Application of MoS2 modified screen-printed electrodes for highly sensitive detection of bovine serum albumin. Anal Chim Acta 939:101–107. https://doi.org/10.1016/j.aca.2016.08.010
Pinwattana K, Wang J, Lin C-T, Wu H, Du D, Lin Y, Chailapakul O (2010) CdSe/ZnS quantum dots based electrochemical immunoassay for the detection of phosphorylated bovine serum albumin. Biosens Bioelectron 26:1109–1113. https://doi.org/10.1016/j.bios.2010.08.021
Guo T, Deng Q, Fang G, Ma L, Wang S (2022) Fluorescence sensor based on molecularly imprinted polymers and core-shell upconversion nanoparticles@metal-organic frameworks for detection of bovine serum albumin. Spectrochim Acta A Mol Biomol Spectrosc 279:121460. https://doi.org/10.1016/j.saa.2022.121460
Gandarilla AMD, Matos RS, Barcelay YR, da Fonseca Filho HD, Brito WR (2022) Molecularly imprinted polymer on indium tin oxide substrate for bovine serum albumin determination. J Polym Res 29:1. https://doi.org/10.1007/s10965-022-03022-5
Yang S, Deng K, Shao S, Zhang J, Peng J, Fang Z, Xu W (2022) A visible light responsive molecularly imprinted photoelectrochemical sensor for the sensitive detection of BSA. J Solid State Electrochem 26:821–830. https://doi.org/10.1007/s10008-021-05110-w
Wei Y, Yu F, Diao Z, Xu R, Li H, Qin G, Guo X (2022) Self-cleaning electrochemical protein-imprinting biosensor with a dual-driven switchable affinity for sensing bovine serum albumin. Talanta 237:122893. https://doi.org/10.1016/j.talanta.2021.122893
Malla P, Liao H-P, Liu C-H, Wu W-C (2021) Electrochemical immunoassay for serum parathyroid hormone using screen-printed carbon electrode and magnetic beads. J Electroanal Chem 895:115463. https://doi.org/10.1016/j.jelechem.2021.115463
Tang X, Catanante G, Huang X, Marty J-L, Wang H, Zhang Q, Li P (2022) Screen-printed electrochemical immunosensor based on a novel nanobody for analyzing aflatoxin M1 in milk. Food Chem 383:132598. https://doi.org/10.1016/j.foodchem.2022.132598
Nelis JLD, Migliorelli D, Mühlebach L, Generelli S, Stewart L, Elliott CT, Campbell K (2021) Highly sensitive electrochemical detection of the marine toxins okadaic acid and domoic acid with carbon black modified screen printed electrodes. Talanta 228:122215. https://doi.org/10.1016/j.talanta.2021.122215
Vukojević V, Djurdjić S, Ognjanović M, Fabián M, Samphao A, Kalcher K, Stanković DM (2018) Enzymatic glucose biosensor based on manganese dioxide nanoparticles decorated on graphene nanoribbons. J Electroanal Chem 823:610–616. https://doi.org/10.1016/j.jelechem.2018.07.013
Stanković V, Đurđić S, Ognjanović M, Mutić J, Kalcher K, Stanković DM (2020) A novel nonenzymatic hydrogen peroxide amperometric sensor based on AgNp@GNR nanocomposites modified screen-printed carbon electrode. J Electroanal Chem 876:114487. https://doi.org/10.1016/j.jelechem.2020.114487
Đurđić S, Vukojević V, Vlahović F, Ognjanović M, Švorc Ľ, Kalcher K, Mutić J, Stanković DM (2019) Application of bismuth (III) oxide decorated graphene nanoribbons for enzymatic glucose biosensing. J Electroanal Chem 850:113400. https://doi.org/10.1016/j.jelechem.2019.113400
Shamsazar A, Asadi A, Seifzadeh D, Mahdavi M (2021) A novel and highly sensitive sandwich-type immunosensor for prostate-specific antigen detection based on MWCNTs-Fe3O4 nanocomposite. Sens Actuators, B Chem 346:130459. https://doi.org/10.1016/j.snb.2021.130459
Viswanathan S, Rani C, J-aA Ho (2012) Electrochemical immunosensor for multiplexed detection of food-borne pathogens using nanocrystal bioconjugates and MWCNT screen-printed electrode. Talanta 94:315–319. https://doi.org/10.1016/j.talanta.2012.03.049
Zhang S, Shen Y, Shen G, Wang S, Shen G, Yu R (2016) Electrochemical immunosensor based on Pd-Au nanoparticles supported on functionalized PDDA-MWCNT nanocomposites for aflatoxin B1 detection. Anal Biochem 494:10–15. https://doi.org/10.1016/j.ab.2015.10.008
Sun X, Li C, Zhu Q, Chen J, Li J, Ding H, Sang F, Kong L, Chen Z, Wei Q (2020) A novel ultrasensitive sandwich-type photoelectrochemical immunoassay for PSA detection based on dual inhibition effect of Au/MWCNTs nanohybrids on N-GQDs/CdS QDs dual sensitized urchin-like TiO2. Electrochim Acta 333:135480. https://doi.org/10.1016/j.electacta.2019.135480
Yola ML, Atar N, Özcan N (2021) A novel electrochemical lung cancer biomarker cytokeratin 19 fragment antigen 21–1 immunosensor based on Si3N4/MoS2 incorporated MWCNTs and core-shell type magnetic nanoparticles. Nanoscale 13:4660–4669. https://doi.org/10.1039/d1nr00244a
Cao L, Tan Y, Deng W, Xie Q (2021) MWCNTs-CoP hybrids for dual-signal electrochemical immunosensing of carcinoembryonic antigen based on overall water splitting. Talanta 233:122521. https://doi.org/10.1016/j.talanta.2021.122521
Liu S, Liu Y, Mu Q, Zhang F, Li H, Wang Y (2013) Synthesis, characterization and photoluminescent properties of rare-earth hydroxides and oxides nanorods by hydrothermal route. Appl Phys A 111:1229–1240. https://doi.org/10.1007/s00339-012-7387-3
Đurđić S, Stanković V, Vlahović F, Ognjanović M, Kalcher K, Manojlović D, Mutić J, Stanković DM (2021) Carboxylated single-wall carbon nanotubes decorated with SiO2 coated-Nd2O3 nanoparticles as an electrochemical sensor for L-DOPA detection. Microchem J 168 106416N. https://doi.org/10.1016/j.microc.2021.106416
Marquez-Mariño K, Penagos-Llanos J, García-Beltrán O, Nagles E, Hurtado JJ (2018) Development of a novel electrochemical sensor based on a carbon paste electrode decorated with Nd 2 O 3 for the simultaneous detection of tartrazine and sunset yellow. Electroanalysis 30:2760–2767. https://doi.org/10.1002/elan.201800550
Nagles E, Calderón JA, García-Beltrán O (2017) Development of an electrochemical sensor to detect dopamine and ascorbic acid based on neodymium (III) oxide and chitosan. Electroanalysis 29:1081–1087. https://doi.org/10.1002/elan.201600729
Arunachalam S, Kirubasankar B, Murugadoss V, Vellasamy D, Angaiah S (2018) Facile synthesis of electrostatically anchored Nd(OH) 3 nanorods onto graphene nanosheets as a high capacitance electrode material for supercapacitors. New J Chem 42:2923–2932. https://doi.org/10.1039/C7NJ04335J
Sanivarapu SR, Lawrence JB, Sreedhar G (2018) Role of surface oxygen vacancies and lanthanide contraction phenomenon of Ln(OH) 3 (Ln = La, Pr, and Nd) in sulfide-mediated photoelectrochemical water splitting. ACS Omega 3:6267–6278. https://doi.org/10.1021/acsomega.8b00429
Qu J, Zheng D, Lu X, Shi J, Lan C (2010) Electrochemical assembling of aligned porous Nd(OH)3 nanobelts with high performance in water treatment. Inorg Chem Commun 13:1425–1428. https://doi.org/10.1016/j.inoche.2010.08.007
Koczkur KM, Mourdikoudis S, Polavarapu L, Skrabalak SE (2015) Polyvinylpyrrolidone (PVP) in nanoparticle synthesis. Dalton Trans 44:17883–17905. https://doi.org/10.1039/C5DT02964C
Heli H, Sattarahmady N, Jabbari A, Moosavi-Movahedi AA, Hakimelahi GH, Tsai F-Y (2007) Adsorption of human serum albumin onto glassy carbon surface – applied to albumin-modified electrode: mode of protein–ligand interactions. J Electroanal Chem 610:67–74. https://doi.org/10.1016/j.jelechem.2007.07.005
Zinatloo-Ajabshir S, Mortazavi-Derazkola S, Salavati-Niasari M (2017) Nd2O3 nanostructures: simple synthesis, characterization and its photocatalytic degradation of methylene blue. J Mol Liq 234:430–436. https://doi.org/10.1016/j.molliq.2017.03.115
Hussein MA, Abu-Zied BM, Asiri AM (2018) Fabrication of EPYR/GNP/MWCNT carbon-based composite materials for promoted epoxy coating performance. RSC Adv 8:23555–23566. https://doi.org/10.1039/c8ra03109f
Chen H-J, Zhang Z-H, Luo L-J, Yao S-Z (2012) Surface-imprinted chitosan-coated magnetic nanoparticles modified multi-walled carbon nanotubes biosensor for detection of bovine serum albumin. Sens Actuators, B Chem 163:76–83. https://doi.org/10.1016/j.snb.2012.01.010
Li M-X, Wang X-H, Zhang L-M, Wei X-P (2017) A high sensitive epitope imprinted electrochemical sensor for bovine serum albumin based on enzyme amplifying. Anal Biochem 530:68–74. https://doi.org/10.1016/j.ab.2017.05.006
Wang L, Ma Y, Wang L (2021) High selectivity sensing of bovine serum albumin: the combination of glass nanopore and molecularly imprinted technology. Biosens Bioelectron 178:113056. https://doi.org/10.1016/j.bios.2021.113056
Yang Y, Pan J, Hua W, Tu Y (2014) An approach for the preparation of highly sensitive electrochemical impedimetric immunosensors for the detection of illicit drugs. J Electroanal Chem 726:1–6. https://doi.org/10.1016/j.jelechem.2014.04.022
Wang Y, Han M, Liu G, Hou X, Huang Y, Wu K, Li C (2015) Molecularly imprinted electrochemical sensing interface based on in-situ-polymerization of amino-functionalized ionic liquid for specific recognition of bovine serum albumin. Biosens Bioelectron 74:792–798. https://doi.org/10.1016/j.bios.2015.07.046
Wei Y, Zeng Q, Hu Q, Wang M, Tao J, Wang L (2018) Self-cleaned electrochemical protein imprinting biosensor basing on a thermo-responsive memory hydrogel. Biosens Bioelectron 99:136–141. https://doi.org/10.1016/j.bios.2017.07.049
Ferreira NS, Moreira APT, de Sá MHM, Sales MGF (2017) New electrochemically-derived plastic antibody on a simple conductive paper support for protein detection: application to BSA. Sens Actuators, B Chem 243:1127–1136. https://doi.org/10.1016/j.snb.2016.12.074
Acknowledgements
The authors gratefully acknowledge the support provided by the Ministry of Education, Science and Technological Development of Republic of Serbia (contract number: 451-03-68/2020-14/200168) and Eureka project E! 13303 MED-BIO-TEST (supported by the Ministry of Education, Science and Technological Development of Republic of Serbia; contract number 451-03-00053/2020-09/2/2).
Author information
Authors and Affiliations
Contributions
Slađana Đurđić: conceptualization, methodology, investigation, writing—original draft. Miloš Ognjanović: investigation, writing, methodology. Maja Krstić Ristivojević: investigation, writing. Bratislav Antić: methodology, validation, writing—original draft. Tanja Ćirković Veličković: methodology, validation, writing—review and editing. Jelena Mutić: methodology, validation. Zoltan Konya: investigation, visualization, writing—review and editing. Dalibor Stanković: conceptualization, visualization, project administration, supervision, writing—review and editing.
Corresponding author
Ethics declarations
Conflict of interest
The authors declare no competing interests.
Additional information
Publisher's note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Information
Below is the link to the electronic supplementary material.
Rights and permissions
Springer Nature or its licensor holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
About this article
Cite this article
Đurđić, S., Ognjanović, M., Ristivojević, M.K. et al. Voltammetric immunoassay based on MWCNTs@Nd(OH)3-BSA-antibody platform for sensitive BSA detection. Microchim Acta 189, 422 (2022). https://doi.org/10.1007/s00604-022-05514-z
Received:
Accepted:
Published:
DOI: https://doi.org/10.1007/s00604-022-05514-z
Keywords
- Bovine serum albumin
- Bovine serum albumin polyclonal antibody
- Electrochemical immunosensor
- Differential pulse voltammetry
- Multi-wall carbon nanotubes
- Neodymium(III) hydroxide