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In veterinary medicine, peptides derived from the immunodominant P44 protein are used to detect antibodies in serological point-of-care tests for anaplasmosis. To inform efforts to develop improved chimeric diagnostic proteins for anaplasmosis, the structure of the predicted surface-exposed domain (residues 151\u2013308) of P44 was determined at a resolution of 1.65 \u00c5. This domain comprises six \u03b1-helices and six short \u03b2-strands connected by loop regions. A structural similarity search of the P44\n                      <jats:sub>151\u2013308<\/jats:sub>\n                      domain using the Distance Matrix Alignment server against the AlphaFold Database revealed that the domain represents a previously uncharacterized protein fold. The determined structure is consistent with AlphaFold v2 predictions, which further predict the core of P44 to be an eight-stranded \u03b2-barrel. Using P44 fragments, immunodominant epitopes were mapped within residues 151\u2013308. Four chimeric genes (ADC1, ADC2, ADC3, and ADC4) were designed that incorporate the analogous immunodominant P44 domains from\n                      <jats:italic toggle=\"yes\">A. phagocytophilum<\/jats:italic>\n                      and\n                      <jats:italic toggle=\"yes\">A. platys<\/jats:italic>\n                      . The recombinant ADC chimerics were assessed for sensitivity and specificity by screening with serum from dogs diagnosed with anaplasmosis, Lyme disease, or ehrlichiosis, with sera from specific pathogen-free dogs serving as negative controls. Accuracy was further demonstrated by screening characterized serum samples from coyotes (\n                      <jats:italic toggle=\"yes\">Canis latrans<\/jats:italic>\n                      ; wild canids). The ADC1 chimeric provided 100% sensitivity and 98.04% specificity. The sensitivity and specificity of ADC1 support its potential use as a diagnostic Ag for anaplasmosis.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1128\/iai.00167-26","type":"journal-article","created":{"date-parts":[[2026,9,15]],"date-time":"2026-09-15T13:40:45Z","timestamp":1789479645000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":0,"title":["Structure-guided design of\n                    <i>Anaplasma<\/i>\n                    P44 protein-based chimeric antigens for serodiagnosis of anaplasmosis in dogs"],"prefix":"10.1128","volume":"94","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1237-4168","authenticated-orcid":false,"given":"Nicholas A.","family":"Cramer","sequence":"first","affiliation":[{"id":[{"id":"https:\/\/ror.org\/057xmsr27","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Microbiology and Immunology, Virginia Commonwealth University Medical 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