TY - JOUR
T1 - Biorecognition and detection of antigens from Mycobacterium tuberculosis using a sandwich ELISA associated with magnetic nanoparticles
AU - León-Janampa, Nancy
AU - Shinkaruk, Svitlana
AU - Gilman, Robert H.
AU - Kirwan, Daniela E.
AU - Fouquet, Eric
AU - Szlosek, Magali
AU - Sheen, Patricia
AU - Zimic, Mirko
N1 - Funding Information:
NLJ thanks the support received by the scholarship of the Franco Peruvian School of Life Sciences (Peru), the faculties and fellow students from Ph.D. program of Life Sciences from Universidad Peruana Cayetano Heredia and University of Bordeaux (France). NLJ received support from ‘Unraveling the resistance mechanism of pyrazinamide, the unique - sterilizing drug against tuberculosis’ (SIDISI: 59921) financed for Wellcome Trust organization. The funders had no role in study design, analysis, decision to publish, or preparation of the manuscript.
Funding Information:
The authors acknowledge Dr M. Gayot of PLACAMAT-Bordeaux, Dr. G. Le Bourdon of the Institute of Molecular Sciences, S. Papillon from Bordeaux Sciences Agro, Adriana del Valle from Universidad Peruana Cayetano Heredia and Katherine Changanaqui from Universidad de Ingeniería (Peru), for collaborating on some measurement techniques applied in this study. NLJ thanks the support received by the scholarship of the Franco Peruvian School of Life Sciences (Peru), the faculties and fellow students from Ph.D. program of Life Sciences from Universidad Peruana Cayetano Heredia and University of Bordeaux (France). NLJ received support from ‘Unraveling the resistance mechanism of pyrazinamide, the unique - sterilizing drug against tuberculosis’ (SIDISI: 59921) financed for Wellcome Trust organization. The funders had no role in study design, analysis, decision to publish, or preparation of the manuscript.
Publisher Copyright:
© 2022 Elsevier B.V.
PY - 2022/6/5
Y1 - 2022/6/5
N2 - Tuberculosis (TB), caused by Mycobacterium tuberculosis (MTB), is one of the 10 leading causes of death worldwide, especially in low-income areas. A rapid, low-cost diagnostic assay for TB with high sensitivity and specificity is not currently available. Bio-functionalized magnetic nanoparticles (MNPs) which are able to efficiently detect and concentrate biomolecules from complex biological samples, allows improving the diagnostic immunoassays. In this way, a proof-of-concept of MNP-based sandwich immunoassay was developed to detect various MTB protein antigens. The superficial and secretory antigenic proteins considered in this research were: CFP10, ESAT6, MTC28, MPT64, 38 kDa protein, Ag85B, and MoeX. The proteins were cloned and expressed in an E. coli system. Polyclonal antibodies (ab) against the recombinant antigens were elicited in rabbits and mice. Antibodies were immobilized on the surface of amine-silanized nanoparticles (MNP@Si). The functionalized MNP@Si@ab were tested in a colorimetric sandwich enzyme-linked immunosorbent assay (sELISA-MNP@Si@ab) to recognize the selected antigens in sputum samples. The selected MTB antigens were successfully detected in sputum from TB patients in a shorter time (~ 4 h) using the sELISA-MNP@Si@ab, compared to the conventional sELISA (~15 h) standardized in home. Moreover, the sELISA-MNP@Si@ab showed the higher sensitivity in the real biological samples from infected patients.
AB - Tuberculosis (TB), caused by Mycobacterium tuberculosis (MTB), is one of the 10 leading causes of death worldwide, especially in low-income areas. A rapid, low-cost diagnostic assay for TB with high sensitivity and specificity is not currently available. Bio-functionalized magnetic nanoparticles (MNPs) which are able to efficiently detect and concentrate biomolecules from complex biological samples, allows improving the diagnostic immunoassays. In this way, a proof-of-concept of MNP-based sandwich immunoassay was developed to detect various MTB protein antigens. The superficial and secretory antigenic proteins considered in this research were: CFP10, ESAT6, MTC28, MPT64, 38 kDa protein, Ag85B, and MoeX. The proteins were cloned and expressed in an E. coli system. Polyclonal antibodies (ab) against the recombinant antigens were elicited in rabbits and mice. Antibodies were immobilized on the surface of amine-silanized nanoparticles (MNP@Si). The functionalized MNP@Si@ab were tested in a colorimetric sandwich enzyme-linked immunosorbent assay (sELISA-MNP@Si@ab) to recognize the selected antigens in sputum samples. The selected MTB antigens were successfully detected in sputum from TB patients in a shorter time (~ 4 h) using the sELISA-MNP@Si@ab, compared to the conventional sELISA (~15 h) standardized in home. Moreover, the sELISA-MNP@Si@ab showed the higher sensitivity in the real biological samples from infected patients.
KW - Active tuberculosis
KW - Biofunctionalization
KW - Immunoassay
KW - Magnetic nanoparticle
KW - Polyclonal antibody
KW - Recombinant protein
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U2 - 10.1016/j.jpba.2022.114749
DO - 10.1016/j.jpba.2022.114749
M3 - Article
C2 - 35447489
AN - SCOPUS:85128279118
SN - 0731-7085
VL - 215
JO - Journal of Pharmaceutical and Biomedical Analysis
JF - Journal of Pharmaceutical and Biomedical Analysis
M1 - 114749
ER -