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Usually, following repeated exposure to pathogens, affinity maturation and clonal selection take place, resulting in increased antibody avidity. However, some studies suggest affinity maturation may not occur to malaria antigens in endemic areas. Information on development of antibody avidity is confusing and conflicting, in part, because different techniques have been used to measure avidity. Today, bead-based multiplex immunoassays (MIA) are routinely used to simultaneously quantitate antibody levels to multiple antigens. This study evaluated the feasibility of developing an avidity MIA with 5 merozoite antigens (AMA1, EBA-175, MSP1-42, MSP2, MSP3) that uses a single chaotropic concentration.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>\n                      The most common ELISA protocols that used the chaotropic reagents guanidine HCl (GdHCl), urea, and ammonium thiocyanate (NH\n                      <jats:sub>4<\/jats:sub>\n                      SCN) were adapted to a multiplex MIA format. Then, different concentrations of chaotropes and incubation times were compared and results were expressed as an Avidity Index (AI), i.e., percentage of antibody remaining bound in the presence of chaotrope. Experiments were conducted to (i) identify the assay with the widest range of AI (discriminatory power), (ii) determine the amount of chaotrope needed to release 50% of bound Ab using plasma from adults and infants, and (iii) evaluate assay repeatability.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      Overall, 4\u00a0M GdHCl and 8\u00a0M urea were weaker chaotropes than 3\u00a0M NH\n                      <jats:sub>4<\/jats:sub>\n                      SCN. For example, they failed to release significant amounts of Ab bound to MSP1-42 in adult plasma samples; whereas, a range of AI values was obtained with NH\n                      <jats:sub>4<\/jats:sub>\n                      SCN. Titration of NH\n                      <jats:sub>4<\/jats:sub>\n                      SCN revealed that 2\u00a0M NH\n                      <jats:sub>4<\/jats:sub>\n                      SCN gave the widest range of AI for the 5 antigens. Binding studies using plasma from 40 adults and 57 1-year old infants in Cameroon showed that 2.1\u00a0M\u2009\u00b1\u20090.32 (mean\u2009\u00b1\u2009SD) NH\n                      <jats:sub>4<\/jats:sub>\n                      SCN (adults) and 1.8\u00a0M\u2009\u00b1\u20090.23\u00a0M (infants) released 50% of bound Ab from the merozoite antigens.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusions<\/jats:title>\n                    <jats:p>\n                      An avidity MIA is feasible for the 5 merozoite antigens that uses a single concentration (2\u00a0M) of NH\n                      <jats:sub>4<\/jats:sub>\n                      SCN. The assay provides a simple method to quickly obtain information about Ab quantity and quality in the acquisition of immunity to malaria in endemic populations.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1186\/s12936-020-03243-3","type":"journal-article","created":{"date-parts":[[2020,5,1]],"date-time":"2020-05-01T14:02:39Z","timestamp":1588341759000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":14,"title":["Measuring antibody avidity to Plasmodium falciparum merozoite antigens using a multiplex immunoassay approach"],"prefix":"10.1186","volume":"19","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-0988-4290","authenticated-orcid":false,"given":"Diane Wallace","family":"Taylor","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Naveen","family":"Bobbili","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alex","family":"Kayatani","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Samuel","family":"Tassi Yunga","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Winifrida","family":"Kidima","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rose F. 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All participants gave written informed consent. The use of deidentified plasma samples for antibody studies was deemed exempt by the Committee on Human Subjects University of Hawai\u2019i (adults: CHS #21891CHS, infants: CHS #21752).","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Ethics approval and consent to participate"}},{"value":"Not applicable.","order":2,"name":"Ethics","group":{"name":"EthicsHeading","label":"Consent for publication"}},{"value":"The authors declare that they have no competing interests.","order":3,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"171"}}