Vol. 12 No. 11 (2026): In Press
Open Access
Peer Reviewed

Development of a Heart Failure Detection Kit Based on Antibody Against Amino-Terminal Pro-Brain Natriuretic Peptide (NT-proBNP): Identification and Immunogenicity Exploration of NT-proBNP Protein

Authors

Andreas Budi Wijaya , Dina Fauziah , Karina Nilasari , Editya Fukata , Dyta Loverita , Taurisma Aulia Nanda Wibisono , Farsya Hidayah Septiananda , Rajendra Aulya Gilardino

DOI:

10.29303/jppipa.v12i11.13398

Published:

2026-11-12

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Abstract

Heart failure (HF) remains a major public health problem with high morbidity and mortality worldwide. The use of cardiac biomarkers such as N-terminal pro–B-type natriuretic peptide (NT-proBNP) has been established as a reliable tool for HF diagnosis and prognosis. However, the high cost and technical requirements of conventional immunoassays limit their accessibility in low-resource settings. This study aims to produce and characterize polyclonal antibodies against NT-proBNP as a preliminary step toward developing an affordable, locally produced detection kit suitable for primary healthcare use in Indonesia. Three male Oryctolagus cuniculus rabbits were immunized intramuscularly with recombinant human NT-proBNP antigen emulsified in Freund’s adjuvant. Immunogenicity was assessed using indirect ELISA at 405 nm absorbance. The ELISA results demonstrated a progressive increase in NT-proBNP-specific antibody titers from week 1 to week 4, with peak levels observed after the second booster (week 7). This pattern reflected the establishment of both primary and secondary humoral immune responses. The absorbance values indicated that NT-proBNP antigen is immunogenic and capable of inducing significant polyclonal antibody production in rabbits. The resulting polyclonal antibodies have multi-epitope recognition potential, which is advantageous for developing point-of-care immunoassays such as lateral flow tests. These results represent an essential foundation for subsequent phases, including antibody purification, affinity testing, and prototype development of an NT-proBNP-based heart failure detection kit optimized for Indonesian healthcare settings. Human NT-proBNP is confirmed to be a highly immunogenic protein capable of inducing the production of specific polyclonal antibodies in rabbits

Keywords:

ELISA Heart failure Immunogenicity Polyclonal antibody NT-proBNP

References

Ali, Z., Hashmi, A., & Gupta, D. (2025). NT-PROBNP As A Prognostic Marker : Correlation Between Biomarker Levels And Severity , Hospitalization Rate , And Mortality In Heart Failure Patient. 14(32), 4602–4609.

Asta, L., Pisano, C., Sbrigata, A., Raffa, G. M., Scola, L., & Balistreri, C. R. (2025). Biomarkers in Heart Failure : A Review and a Wish. LV, 1–35.

Chowdhury, R. R., Kaur, S., & Gera, R. (2023). N-Terminal Pro-B-Type Natriuretic Peptide as a Marker of Severity of Heart Failure in Children with Congenital Heart Diseases. Pediatric Cardiology, 44(8), 1716–1720. https://doi.org/10.1007/S00246-023-03259-X

Clerico, A., Giannoni, A., Vittorini, S., & Passino, C. (2011). Thirty years of the heart as an endocrine organ : physiological role and clinical utility of cardiac natriuretic hormones. 12–20. https://doi.org/10.1152/ajpheart.00226.2011.

Daniels, L. B., Maisel, A. S., & Diego, S. (2007). Natriuretic Peptides. 50(25). https://doi.org/10.1016/j.jacc.2007.09.021

Deluca, K. F., Mick, J. E., & Deluca, J. G. (2022). Protocol Production and purification of recombinant monoclonal antibodies from human cells based on a primary sequence Production and purification of recombinant monoclonal antibodies from human cells based on a primary. STAR Protocols, 3(4), 101915. https://doi.org/10.1016/j.xpro.2022.101915

Internal, E., Clinic, M., & Merkur, C. H. (2023). function in cardiorenal syndromes : a narrative review. 33(3), 1–13.

Lee, Y., Choi, D., & Kim, G. (2021). Development of a Rapid Diagnostic Kit for Congestive Heart Failure Using Recombinant NT-proBNP Antigen.

Li, H., Yin, X., Sun, D., Xia, K., Kang, C., Chu, S., Zhang, P., Wang, H., & Qiu, Y. (2016). Sensing and Bio-Sensing Research Detection of NT-pro BNP using fl uorescent protein modi fi ed by streptavidin as a label in immunochromatographic assay. SBSR, 11, 1–7. https://doi.org/10.1016/j.sbsr.2016.08.001

Posthuma-trumpie, G. A., & Korf, J. (2009). Lateral flow ( immuno ) assay : its strengths , weaknesses , opportunities and threats . A literature survey. 569–582. https://doi.org/10.1007/s00216-008-2287-2

Reeves, S., Womack, C., Lutherer, L., Todd, C., Pinkney, K., & Kasemsri, T. (2018). What Is High Enough? Elevated NT-pro-BNP in Decompensated Paroxysmal Supraventricular Tachycardia. Journal of Pediatric Intensive Care, 7(1), 049–053. https://doi.org/10.1055/S-0037-1603760

Tarmizi, M., Jared, S., Yang, Y., Nikki, U. B., Tan, Y., Wan, Y., Luke, B., & Chamindie, S. (2014). ORIGINAL RESEARCH PAPER A rapid and cost-effective method of producing recombinant proBNP and NT-proBNP variants in Escherichia coli for immunoassay of heart failure. 133–140. https://doi.org/10.1007/s10529-013-1341-0

Yancy, C. W., Jessup, M., Bozkurt, B., Butler, J., Casey, D. E., Drazner, M. H., Fonarow, G. C., Geraci, S. A., Horwich, T., Januzzi, J. L., Johnson, M. R., Kasper, E. K., Levy, W. C., Masoudi, F. A., McBride, P. E., McMurray, J. J. V., Mitchell, J. E., Peterson, P. N., Riegel, B., … Wilkoff, B. L. (2013). 2013 ACCF/AHA guideline for the management of heart failure: A report of the american college of cardiology foundation/american heart association task force on practice guidelines. Circulation, 128(16), 240–327. https://doi.org/10.1161/CIR.0b013e31829e8776

Zhang, X., Zhai, Z., Geng, X., Zhu, Y., Yang, S., Chen, T., Wu, X., Zhang, J., Guo, Z. G., & Hou, M. (2023). Doppler Echocardiography Combined with NTproBNP/BNP in the Diagnosis of Pulmonary Artery Hypertension Associated with Congenital Heart Disease. Computational and Mathematical Methods in Medicine, 2023, 1–5. https://doi.org/10.1155/2023/1896026

Author Biographies

Andreas Budi Wijaya, Universitas Negeri Malang

Author Origin : Indonesia

Dina Fauziah, Universitas Negeri Malang

Author Origin : Indonesia

Karina Nilasari, Universitas Negeri Malang

Author Origin : Indonesia

Editya Fukata, Universitas Negeri Malang

Author Origin : Indonesia

Dyta Loverita, Universitas Negeri Malang

Author Origin : Indonesia

Taurisma Aulia Nanda Wibisono, Universitas Negeri Malang

Author Origin : Indonesia

Farsya Hidayah Septiananda, Universitas Negeri Malang

Author Origin : Indonesia

Rajendra Aulya Gilardino, Universitas Negeri Malang

Author Origin : Indonesia

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How to Cite

Wijaya, A. B., Fauziah, D., Nilasari, K., Fukata, E., Loverita, D., Wibisono, T. A. N., … Gilardino, R. A. (2026). Development of a Heart Failure Detection Kit Based on Antibody Against Amino-Terminal Pro-Brain Natriuretic Peptide (NT-proBNP): Identification and Immunogenicity Exploration of NT-proBNP Protein. Jurnal Penelitian Pendidikan IPA, 12(11), 36–41. https://doi.org/10.29303/jppipa.v12i11.13398