Evaluation of anti-histidine-rich protein 2 monoclonal antibodies, developed by using poly (N-isopropylacrylamide) as an adjuvant for malarial diagnostic application
https://doi.org/10.47665/tb.37.4.1050
- Author:
Vishalkumar, P.
1
;
Jayaprakash, N.S.
2
;
Desai, P.K.
1
;
Krishnan, V.
2
;
Vijayalakshmi, M.A.
2
Author Information
1. Arkray Healthcare Pvt Ltd, Plot No. 336, 338, 340, Rd Number 30, Sachin 394230, Surat, India
2. Centre for Bioseparation Technology (CBST), Vellore Institute of Technology (VIT), Vellore 632014, Tamil Nadu, India
- Publication Type:Journal Article
- From:Tropical Biomedicine
2020;37(No.4):1050-1061
- CountryMalaysia
- Language:English
-
Abstract:
Objective: To evaluate the sensitivity and the stability of the monoclonal antibodies
(Aa3c10, b10c1), against truncated Histidine-rich protein 2 (PfHRP2), developed using smart
polymer, poly N-isopropylacrylamide, as adjuvant for malarial diagnostic applications in
comparison with the available commercial antibodies. Methods: Two hybridoma clones (Aa3c10,
b10c1) were used for the production of ascites in BALB/c mice. Purification of monoclonal
antibodies from the ascites was carried out using affinity columns. The thermal stability
study of monoclonal antibodies was done by storing it at 37°C and 45°C for thirty days. The
stored antibodies were analyzed using SDS-PAGE and flow-through device where the antigenantibody interaction was visualized by Protein A colloidal gold solution. Sensitivity was
determined by endpoint dilution ELISA and the dissociation constant by competitive ELISA.
Sensitive pair optimization was done by sandwich ELISA using biotinylated antibodies.
Prototype preparation for lateral flow assay had a colloidal gold-based detection system.
Results: Thermal stability experiments showed that both mAbs (Aa3c10; b10c1) are stable up
to thirty days at 45°C while the commercially available mAbs were stable up to fifteen days
only. Compared to commercial antibodies, the mAb Aa3c10, showed the highest sensitivity in
end-point titre. In sensitive pair optimization, it was observed that the mAb, b10c1, as a
detector and the mAb, Aa3c10, as a capture antibody showed the highest absorbance to
detect 50pg/ml PfHRP2 antigen. The prototype formulation of lateral flow assay using the
mAbs (Aa3c10; b10c1) showed good reactivity with WHO panel and no false-positive results
were observed with twenty clinically negative samples and five P. vivax positive samples.
Conclusions: The novel monoclonal antibodies (Aa3c10, b10c1) against truncated PfHRP2,
could be a strong potential candidates that can be included in making RDTs with better
sensitivity and stability.
- Full text:8.2020my1189.pdf
