Evaluation of 1st WHO Anti-Malaria Reference Reagent for Competition ELISA Harmonisation and Development of ADAMSEL Analytical Platform

This study focuses on harmonising the competition ELISA (cELISA) assay for Plasmodium falciparum ( P. falciparum ), using the 1st WHO reference reagent for anti-malaria ( P. falciparum ) human reference serum (10/198). Antibody-mediated immune responses, against the Apical Membrane Antigen 1 (AMA1), play a significant role in protection against malaria. However, the sequence diversity of AMA1 and cross-reactivity among variants pose challenges in assessing antibody responses. To address this, the cELISA assay was selected to examine cross-reactive antibody responses against different variants. The harmonisation process for cELISA was performed in three laboratories. The 10/198 served as an internal standard for the calculation of IgG concentrations in the cELISA using ADAMSEL software. Additionally, a novel semi-automated analytical tool was developed in the R-statistics environment. This tool is freely available for download and streamlines result in generation while minimising human error. This study demonstrated the effectiveness of the 1st WHO reference reagent as a standard for ELISA. Additionally, the ADAMSEL software and R-platform tool provide a user-friendly and accessible tool for the analysis of cELISA data. Its automation capabilities improve efficiency and ensure global accessibility at no cost, benefitting laboratories with limited resources. Importance This study focuses on the 1st WHO reference reagent (10/198) in harmonizing the competition ELISA (cELISA) assay for P. falciparum. In addition, the introduction of ADAMSEL software for cELISA data analysis, coupled with a novel R-platform tool, available for free, simplifies analytical processes and contributes to global accessibility. This study demonstrates the 1st WHO reference reagent’s efficacy as a competition ELISA standard. ### Competing Interest Statement The authors have declared no competing interest. ### Funding Statement This study was funded by the FP7 EURIPRED (FP7-INFRA-2012 Grant Agreement No. 312661) ### Author Declarations I confirm all relevant ethical guidelines have been followed, and any necessary IRB and/or ethics committee approvals have been obtained. Yes The details of the IRB/oversight body that provided approval or exemption for the research described are given below: N/A I confirm that all necessary patient/participant consent has been obtained and the appropriate institutional forms have been archived, and that any patient/participant/sample identifiers included were not known to anyone (e.g., hospital staff, patients or participants themselves) outside the research group so cannot be used to identify individuals. Yes I understand that all clinical trials and any other prospective interventional studies must be registered with an ICMJE-approved registry, such as ClinicalTrials.gov. I confirm that any such study reported in the manuscript has been registered and the trial registration ID is provided (note: if posting a prospective study registered retrospectively, please provide a statement in the trial ID field explaining why the study was not registered in advance). Yes I have followed all appropriate research reporting guidelines, such as any relevant EQUATOR Network research reporting checklist(s) and other pertinent material, if applicable. Yes All data produced in the present work are contained in the manuscript