{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"Research Square"}],"indexed":{"date-parts":[[2026,7,13]],"date-time":"2026-07-13T17:04:30Z","timestamp":1783962270439,"version":"3.55.0"},"posted":{"date-parts":[[2025,6,21]]},"group-title":"In Review","reference-count":39,"publisher":"Springer Science and Business Media LLC","license":[{"start":{"date-parts":[[2025,6,21]],"date-time":"2025-06-21T00:00:00Z","timestamp":1750464000000},"content-version":"unspecified","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2025,5,1]]},"abstract":"<title>Abstract<\/title>\n                <p>\n                  This study was conducted to yield a robust and scalable manufacturing process for a candidate vaccine for human lymphatic filariasis (LF) - a tropical parasitic infection transmitted by mosquitoes. In previous studies, we demonstrated that removing an affinity purification tag from the fusion protein did not affect immunogenicity or vaccine efficacy. During scaled-up production of r\n                  <italic>Bm<\/italic>\n                  HAXT, we noticed that significant amounts of the antigen aggregated, resulting in the loss of purified vaccine antigens. Thus, this project aimed to create new r\n                  <italic>Bm<\/italic>\n                  HAXT forms more suitable for industrial-scale production while maintaining robust protection. We generated three different variants: one with all the cysteinyl residues mutated to serinyl residues (delta-Cys), a second one with a flexible glycine-serine linker inserted between each of the component antigens (GS), and finally, a third variant with a combination of both the cysteine deletion and the addition of linkers (delta-Cys GS). We then evaluated the immunogenicity and efficacy of each variant in a mouse model. We demonstrated that the delta-Cys mutant retained immunogenicity and vaccine efficacy similar to the parent tag-free r\n                  <italic>Bm<\/italic>\n                  HAXT protein. We also evaluated the proteins in an accelerated stability study at five (5) different temperatures (-80\u00b0C, -20\u00b0C, 4\u00b0C, 25\u00b0C, and 40\u00b0C). We concluded that all preparations were stable at 4\u00b0C, and the delta-Cys variant was stable even at 25\u00b0C up to the completion of the study (6 weeks). In addition to improved stability, the delta-Cys protein exhibited reduced aggregation and equivalent potency in mice and, therefore, is an optimal candidate for progression to cGMP manufacturing and human clinical trials as a vaccine for lymphatic filariasis.\n                <\/p>","DOI":"10.21203\/rs.3.rs-6572437\/v1","type":"posted-content","created":{"date-parts":[[2025,6,21]],"date-time":"2025-06-21T01:33:43Z","timestamp":1750469623000},"source":"Crossref","is-referenced-by-count":0,"title":["A Next-Generation Human Lymphatic Filariasis Vaccine Candidate, rBmHAXT, for Clinical Development"],"prefix":"10.21203","author":[{"given":"Nithila","family":"Saravanan","sequence":"first","affiliation":[{"name":"University of Illinois at Rockford"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sean","family":"Gray","sequence":"additional","affiliation":[{"name":"PAI Life Sciences, Inc"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jennifer","family":"Davis","sequence":"additional","affiliation":[{"name":"PAI Life Sciences, Inc"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Conrad M","family":"Puff-Carter","sequence":"additional","affiliation":[{"name":"PAI Life Sciences, Inc"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Vishal","family":"Khatri","sequence":"additional","affiliation":[{"name":"University of Illinois at Rockford"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nikhil","family":"Chauhan","sequence":"additional","affiliation":[{"name":"University of Illinois at Rockford"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Darrick","family":"Carter","sequence":"additional","affiliation":[{"name":"PAI Life Sciences, Inc"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ramaswamy","family":"Kalyanasundaram","sequence":"additional","affiliation":[{"name":"University of Illinois at Rockford"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","reference":[{"key":"ref1","unstructured":"World Health Organization (WHO, https:\/\/www.who.int\/news-room\/fact-sheets\/detail\/lymphatic-filariasis"},{"key":"ref2","doi-asserted-by":"publisher","first-page":"E44","DOI":"10.3390\/tropicalmed4010044","article-title":"The Global Burden of Disease of Zoonotic Parasitic Diseases: Top 5 Contenders for Priority Consideration","volume":"4","author":"Pisarski K","year":"2019","unstructured":"Pisarski, K. 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