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Evaluation of the precision of the Plasmodium knowlesi growth inhibition assay for Plasmodium vivax Duffy-binding protein-based malaria vaccine development
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Evaluation of the precision of the Plasmodium knowlesi growth inhibition assay for Plasmodium vivax Duffy-binding protein-based malaria vaccine development
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Evaluation of the precision of the Plasmodium knowlesi growth inhibition assay for Plasmodium vivax Duffy-binding protein-based malaria vaccine development
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Journal Article
Evaluation of the precision of the Plasmodium knowlesi growth inhibition assay for Plasmodium vivax Duffy-binding protein-based malaria vaccine development
2024
Overview
•The PkGIA will be an essential selection tool for P. vivax vaccine development.•The error of the assay was evaluated with human anti-PvDBPII antibodies.•Significant assay-to-assay variation was observed.•95 % confidence interval of inhibition for a given number of PkGIA was determined.
Recent data indicate increasing disease burden and importance of Plasmodium vivax (Pv) malaria. A robust assay will be essential for blood-stage Pv vaccine development. Results of the in vitro growth inhibition assay (GIA) with transgenic P. knowlesi (Pk) parasites expressing the Pv Duffy-binding protein region II (PvDBPII) correlate with in vivo protection in the first PvDBPII controlled human malaria infection (CHMI) trials, making the PkGIA an ideal selection tool once the precision of the assay is defined. To determine the precision in percentage of inhibition in GIA (%GIA) and in GIA50 (antibody concentration that gave 50 %GIA), ten GIAs with transgenic Pk parasites were conducted with four different anti-PvDBPII human monoclonal antibodies (mAbs) at concentrations of 0.016 to 2 mg/mL, and three GIAs with eighty anti-PvDBPII human polyclonal antibodies (pAbs) at 10 mg/mL. A significant assay-to-assay variation was observed, and the analysis revealed a standard deviation (SD) of 13.1 in the mAb and 5.94 in the pAb dataset for %GIA, with a LogGIA50 SD of 0.299 (for mAbs). Moreover, the ninety-five percent confidence interval (95 %CI) for %GIA or GIA50 in repeat assays was calculated in this investigation. The error range determined in this study will help researchers to compare PkGIA results from different assays and studies appropriately, thus supporting the development of future blood-stage malaria vaccine candidates, specifically second-generation PvDBPII-based formulations.
Publisher
Elsevier Ltd,Elsevier Limited
Subject
/ Animals
/ Antibodies, Monoclonal - immunology
/ Antibodies, Protozoan - blood
/ Antibodies, Protozoan - immunology
/ Antigens
/ Antigens, Protozoan - genetics
/ Antigens, Protozoan - immunology
/ Assaying
/ Blood
/ Cloning
/ genetically modified organisms
/ Genomes
/ Humans
/ Malaria
/ Malaria Vaccines - immunology
/ Malaria, Vivax - prevention & control
/ Plasmodium knowlesi - genetics
/ Plasmodium knowlesi - immunology
/ Plasmodium vivax - immunology
/ Proteins
/ Protozoa
/ Protozoan Proteins - genetics
/ Protozoan Proteins - immunology
/ PvDBPII
/ Receptors, Cell Surface - genetics
/ Receptors, Cell Surface - immunology
/ Vaccine
/ Vaccine Development - methods
/ Vaccines
