{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"Research Square"}],"indexed":{"date-parts":[[2024,1,26]],"date-time":"2024-01-26T00:10:39Z","timestamp":1706227839231},"posted":{"date-parts":[[2023,12,19]]},"group-title":"In Review","reference-count":41,"publisher":"Research Square Platform LLC","license":[{"start":{"date-parts":[[2023,12,19]],"date-time":"2023-12-19T00:00:00Z","timestamp":1702944000000},"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":[[2023,12,15]]},"abstract":"<jats:title>Abstract<\/jats:title>\n        <jats:p>Vector control is a crucial strategy for malaria elimination by preventing infection and reducing disease transmission. Most gains have been achieved through insecticide-treated nets (ITNs) and indoor residual spraying (IRS), but the emergence of insecticide resistance among <jats:italic>Anopheles<\/jats:italic> mosquitoes calls for new tools to be applied. Here, we present the development of a highly effective murine monoclonal antibody, targeting the N-terminal region of the <jats:italic>Plasmodium falciparum<\/jats:italic> gametocyte antigen Pfs230, that can decrease the infection prevalence by &gt;\u200950% when fed to <jats:italic>Anopheles<\/jats:italic> mosquitoes with gametocytes in an artificial membrane feeding system. We used a standard mouse immunization protocol followed by protein interaction and parasite-blocking validation at three distinct stages of the monoclonal antibody development pipeline: post-immunization, post-hybridoma generation, and final validation of the monoclonal antibody. We evaluated twenty antibodies identifying one (mAb 13G9) with high Pfs230-affinity and parasite-blocking activity. This 13G9 monoclonal antibody could potentially be developed into a transmission-blocking single-chain antibody for expression in transgenic mosquitoes.<\/jats:p>","DOI":"10.21203\/rs.3.rs-3757253\/v1","type":"posted-content","created":{"date-parts":[[2023,12,19]],"date-time":"2023-12-19T20:44:53Z","timestamp":1703018693000},"source":"Crossref","is-referenced-by-count":0,"title":["Development of an anti-Pfs230 monoclonal antibody as a Plasmodium falciparum gametocyte blocker"],"prefix":"10.21203","author":[{"given":"Emilia C.","family":"Cuccurullo","sequence":"first","affiliation":[{"name":"Johns Hopkins University"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yuemei","family":"Dong","sequence":"additional","affiliation":[{"name":"Johns Hopkins University"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Maria L.","family":"Sim\u00f5es","sequence":"additional","affiliation":[{"name":"Johns Hopkins University"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"George","family":"Dimopoulos","sequence":"additional","affiliation":[{"name":"Johns Hopkins University"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ethan","family":"Bier","sequence":"additional","affiliation":[{"name":"University of California, San Diego"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"8761","reference":[{"key":"ref1","unstructured":"World Health Organization. World malaria report 2022 [Internet]. Available from: https:\/\/www.who.int\/teams\/global-malaria-programme."},{"key":"ref2","unstructured":"World Health Organization. World malaria report 2020\u201320 years of global progress & challenges."},{"issue":"11","key":"ref3","doi-asserted-by":"crossref","first-page":"1005","DOI":"10.1056\/NEJMoa2026330","article-title":"Seasonal Malaria Vaccination with or without Seasonal Malaria Chemoprevention","volume":"385","author":"Chandramohan D","year":"2021","unstructured":"Chandramohan D, Zongo I, Sagara I, Cairns M, Yerbanga RS, Diarra M, et al. Seasonal Malaria Vaccination with or without Seasonal Malaria Chemoprevention. N Engl J Med. 2021;385(11):1005\u201317.","journal-title":"N Engl J Med"},{"key":"ref4","volume-title":"Plasmodium falciparum gametocytes: Playing hide and seek. Vol.\u00a03, Annals of Translational Medicine","author":"Gardiner DL","year":"2015","unstructured":"Gardiner DL, Trenholme KR. Plasmodium falciparum gametocytes: Playing hide and seek. Vol.\u00a03, Annals of Translational Medicine. AME Publishing Company; 2015."},{"key":"ref5","first-page":"177","volume-title":"Plasmodium asexual growth and sexual development in the haematopoietic niche of the host","author":"Venugopal K","year":"2020","unstructured":"Venugopal K, Hentzschel F, Valki\u016bnas G, Marti M. Plasmodium asexual growth and sexual development in the haematopoietic niche of the host. Nature Reviews Microbiology. Volume 18. Nature Research; 2020. pp. 177\u201389."},{"issue":"3","key":"ref6","doi-asserted-by":"crossref","DOI":"10.1038\/73062","article-title":"Malaria transmission-blocking vaccines - how can their development be supported","volume":"6","author":"Carter R","year":"2000","unstructured":"Carter R, Mendis KN, Miller LH, Molineaux L, Saul A. Malaria transmission-blocking vaccines - how can their development be supported. Nat Med. 2000;6(3).","journal-title":"Nat Med"},{"key":"ref7","author":"Carter R","year":"2001","unstructured":"Carter R. Transmission blocking malaria vaccines. Vaccine [Internet]. 2001;19(17):2309\u201314. Available from: https:\/\/www.sciencedirect.com\/science\/article\/pii\/S0264410X00005211."},{"key":"ref8","first-page":"54","volume-title":"Mosquito transgenesis for malaria control","author":"Dong S","year":"2022","unstructured":"Dong S, Dong Y, Sim\u00f5es ML, Dimopoulos G. Mosquito transgenesis for malaria control. Trends in Parasitology. Volume 38. Elsevier Ltd; 2022. pp. 54\u201366."},{"key":"ref9","author":"Barr PJ","unstructured":"Barr PJ, Green KM, Gibson HL, Bathurst IC, Quakyi IA, Kaslow DC. Recombinant Pfs25 Protein of Plasmoch\u2019um falciparum Elicits Malaria Transmission-blocking Immunity in Experimental Animals [Internet]. Available from: http:\/\/rupress.org\/jem\/article-pdf\/174\/5\/1203\/1672511\/1203.pdf."},{"issue":"1","key":"ref10","doi-asserted-by":"crossref","DOI":"10.1038\/s41467-019-11980-6","article-title":"Potent antibody lineage against malaria transmission elicited by human vaccination with Pfs25","volume":"10","author":"McLeod B","year":"2019","unstructured":"McLeod B, Miura K, Scally SW, Bosch A, Nguyen N, Shin H et al. Potent antibody lineage against malaria transmission elicited by human vaccination with Pfs25. Nat Commun. 2019;10(1).","journal-title":"Nat Commun"},{"issue":"1","key":"ref11","doi-asserted-by":"crossref","DOI":"10.1038\/s41467-017-01924-3","article-title":"Molecular definition of multiple sites of antibody inhibition of malaria transmission-blocking vaccine antigen Pfs25","volume":"8","author":"Scally SW","year":"2017","unstructured":"Scally SW, McLeod B, Bosch A, Miura K, Liang Q, Carroll S et al. Molecular definition of multiple sites of antibody inhibition of malaria transmission-blocking vaccine antigen Pfs25. Nat Commun. 2017;8(1).","journal-title":"Nat Commun"},{"issue":"2","key":"ref12","doi-asserted-by":"crossref","first-page":"161","DOI":"10.1111\/j.1365-3024.1991.tb00272.x","article-title":"Fine specificities of monoclonal antibodies against the Plasmodium falciparum circumsporozoite protein: recognition of both repetitive and non-repetitive regions","volume":"13","author":"Burkot TR","year":"1991","unstructured":"Burkot TR, Da ZW, Geysen HM, Wirtz RA, Saul A. Fine specificities of monoclonal antibodies against the Plasmodium falciparum circumsporozoite protein: recognition of both repetitive and non-repetitive regions. Parasite Immunol. 1991;13(2):161\u201370.","journal-title":"Parasite Immunol"},{"issue":"6","key":"ref13","doi-asserted-by":"crossref","DOI":"10.15252\/emmm.202317454","article-title":"Molecular and functional properties of human Plasmodium falciparum CSP C-terminus antibodies","volume":"15","author":"Oludada OE","year":"2023","unstructured":"Oludada OE, Costa G, Burn Aschner C, Obraztsova AS, Prieto K, Canetta C et al. Molecular and functional properties of human Plasmodium falciparum CSP C-terminus antibodies. EMBO Mol Med. 2023;15(6).","journal-title":"EMBO Mol Med"},{"key":"ref14","first-page":"329","volume-title":"Towards clinical development of a Pfs48\/45-based transmission blocking malaria vaccine","author":"Theisen M","year":"2017","unstructured":"Theisen M, Jore MM, Sauerwein R. Towards clinical development of a Pfs48\/45-based transmission blocking malaria vaccine. Expert Review of Vaccines. Volume 16. Taylor and Francis Ltd; 2017. pp. 329\u201336."},{"issue":"1","key":"ref15","doi-asserted-by":"crossref","DOI":"10.1038\/s41467-018-06742-9","article-title":"Structural delineation of potent transmission-blocking epitope I on malaria antigen Pfs48\/45","volume":"9","author":"Kundu P","year":"2018","unstructured":"Kundu P, Semesi A, Jore MM, Morin MJ, Price VL, Liang A et al. Structural delineation of potent transmission-blocking epitope I on malaria antigen Pfs48\/45. Nat Commun. 2018;9(1).","journal-title":"Nat Commun"},{"issue":"1","key":"ref16","doi-asserted-by":"crossref","DOI":"10.1038\/s41467-021-21955-1","article-title":"A human monoclonal antibody blocks malaria transmission and defines a highly conserved neutralizing epitope on gametes","volume":"12","author":"Coelho CH","year":"2021","unstructured":"Coelho CH, Tang WK, Burkhardt M, Galson JD, Muratova O, Salinas ND et al. A human monoclonal antibody blocks malaria transmission and defines a highly conserved neutralizing epitope on gametes. Nat Commun. 2021;12(1).","journal-title":"Nat Commun"},{"issue":"1","key":"ref17","doi-asserted-by":"crossref","DOI":"10.1038\/s41541-018-0065-5","article-title":"Antibody targeting of a specific region of Pfs47 blocks Plasmodium falciparum malaria transmission","volume":"3","author":"Canepa GE","year":"2018","unstructured":"Canepa GE, Molina-Cruz A, Yenkoidiok-Douti L, Calvo E, Williams AE, Burkhardt M et al. Antibody targeting of a specific region of Pfs47 blocks Plasmodium falciparum malaria transmission. NPJ Vaccines. 2018;3(1).","journal-title":"NPJ Vaccines"},{"issue":"1","key":"ref18","doi-asserted-by":"crossref","DOI":"10.1038\/s41598-019-53208-z","article-title":"Engineering a Virus-Like Particle as an Antigenic Platform for a Pfs47-Targeted Malaria Transmission-Blocking Vaccine","volume":"9","author":"Yenkoidiok-Douti L","year":"2019","unstructured":"Yenkoidiok-Douti L, Williams AE, Canepa GE, Molina-Cruz A, Barillas-Mury C. Engineering a Virus-Like Particle as an Antigenic Platform for a Pfs47-Targeted Malaria Transmission-Blocking Vaccine. Sci Rep. 2019;9(1).","journal-title":"Sci Rep"},{"key":"ref19","author":"Dijk MR","year":"2001","unstructured":"Van Dijk MR, Janse CJ, Thompson J, Waters AP, Braks JAM, Dodemont HJ et al. A Central Role for P48\/45 in Malaria Parasite Male Gamete Fertility The central role of zygote formation in the life cycle and transmission of the parasite makes gametes and. Vol.\u00a0104, Cell. 2001."},{"issue":"5","key":"ref20","doi-asserted-by":"crossref","first-page":"675","DOI":"10.1016\/j.cell.2005.03.027","article-title":"Proteome analysis of separated male and female gametocytes reveals novel sex-specific Plasmodium biology","volume":"121","author":"Khan SM","year":"2005","unstructured":"Khan SM, Franke-Fayard B, Mair GR, Lasonder E, Janse CJ, Mann M, et al. Proteome analysis of separated male and female gametocytes reveals novel sex-specific Plasmodium biology. Cell. 2005;121(5):675\u201387.","journal-title":"Cell"},{"issue":"2","key":"ref21","doi-asserted-by":"crossref","first-page":"216","DOI":"10.1016\/j.molbiopara.2006.05.015","article-title":"Pfs47, paralog of the male fertility factor Pfs48\/45, is a female specific surface protein in Plasmodium falciparum","volume":"149","author":"Schaijk BCL","year":"2006","unstructured":"van Schaijk BCL, van Dijk MR, van de Vegte-Bolmer M, van Gemert GJ, van Dooren MW, Eksi S, et al. Pfs47, paralog of the male fertility factor Pfs48\/45, is a female specific surface protein in Plasmodium falciparum. Mol Biochem Parasitol. 2006;149(2):216\u201322.","journal-title":"Mol Biochem Parasitol"},{"issue":"4","key":"ref22","doi-asserted-by":"crossref","first-page":"991","DOI":"10.1111\/j.1365-2958.2006.05284.x","article-title":"Malaria transmission-blocking antigen, Pfs230, mediates human red blood cell binding to exflagellating male parasites and oocyst production","volume":"61","author":"Eksi S","year":"2006","unstructured":"Eksi S, Czesny B, Van Gemert GJ, Sauerwein RW, Eling W, Williamson KC. Malaria transmission-blocking antigen, Pfs230, mediates human red blood cell binding to exflagellating male parasites and oocyst production. Mol Microbiol. 2006;61(4):991\u20138.","journal-title":"Mol Microbiol"},{"issue":"38","key":"ref23","doi-asserted-by":"crossref","first-page":"19913","DOI":"10.1074\/jbc.M116.732305","article-title":"Structural and immunological characterization of recombinant 6-cysteine domains of the plasmodium falciparum sexual stage protein Pfs230","volume":"291","author":"MacDonald NJ","year":"2016","unstructured":"MacDonald NJ, Nguyen V, Shimp R, Reiter K, Herrera R, Burkhardt M, et al. Structural and immunological characterization of recombinant 6-cysteine domains of the plasmodium falciparum sexual stage protein Pfs230. J Biol Chem. 2016;291(38):19913\u201322.","journal-title":"J Biol Chem"},{"key":"ref24","author":"Brooks SR","year":"2000","unstructured":"Brooks SR, Williamson KC. Proteolysis of Plasmodium falciparum surface antigen, Pfs230, during gametogenesis [Internet]. Vol.\u00a0106, Molecular and Biochemical Parasitology. 2000. Available from: www.elsevier.com\/locate\/parasitology."},{"issue":"1","key":"ref25","doi-asserted-by":"crossref","first-page":"33","DOI":"10.1016\/0166-6851(95)02507-3","article-title":"Recombinant Pfs230, a Plasmodium falciparum gametocyte protein, induces antisera that reduce the infectivity of Plasmodium falciparum to mosquitoes","volume":"75","author":"Williamson KC","year":"1995","unstructured":"Williamson KC, Keister DB, Muratova O, Kaslow DC. Recombinant Pfs230, a Plasmodium falciparum gametocyte protein, induces antisera that reduce the infectivity of Plasmodium falciparum to mosquitoes. Mol Biochem Parasitol. 1995;75(1):33\u201342.","journal-title":"Mol Biochem Parasitol"},{"issue":"1","key":"ref26","doi-asserted-by":"crossref","DOI":"10.1038\/s42003-020-01123-9","article-title":"Structure and function of a malaria transmission blocking vaccine targeting Pfs230 and Pfs230-Pfs48\/45 proteins","volume":"3","author":"Singh K","year":"2020","unstructured":"Singh K, Burkhardt M, Nakuchima S, Herrera R, Muratova O, Gittis AG et al. Structure and function of a malaria transmission blocking vaccine targeting Pfs230 and Pfs230-Pfs48\/45 proteins. Commun Biol. 2020;3(1).","journal-title":"Commun Biol"},{"key":"ref27","article-title":"Transmission-blocking antibodies against multiple, non-variant target epitopes of the Plasmodium fakiparum gamete surface antigen Pfs230 are all complement-fixing","author":"Read D","year":"1994","unstructured":"Read D, Lensen AH, Begarnie S, Haley S, Raza A, Carter R. Transmission-blocking antibodies against multiple, non-variant target epitopes of the Plasmodium fakiparum gamete surface antigen Pfs230 are all complement-fixing. Parasite Immunol. 1994.","journal-title":"Parasite Immunol"},{"issue":"12","key":"ref28","doi-asserted-by":"crossref","first-page":"4377","DOI":"10.1128\/IAI.01056-13","article-title":"Functional comparison of plasmodium falciparum transmission-blocking vaccine candidates by the standard membrane-feeding assay","volume":"81","author":"Miura K","year":"2013","unstructured":"Miura K, Takashima E, Deng B, Tullo G, Diouf A, Moretz SE, et al. Functional comparison of plasmodium falciparum transmission-blocking vaccine candidates by the standard membrane-feeding assay. Infect Immun. 2013;81(12):4377\u201382.","journal-title":"Infect Immun"},{"key":"ref29","first-page":"119","article-title":"Generation of murine monoclonal antibodies by hybridoma technology","volume":"2017","author":"Holzl\u00f6hner P","year":"2017","unstructured":"Holzl\u00f6hner P, Hanack K. Generation of murine monoclonal antibodies by hybridoma technology. J Visualized Experiments. 2017;2017:119.","journal-title":"J Visualized Experiments"},{"key":"ref30","first-page":"653","volume-title":"Toward the development of effective transmission-blocking vaccines for malaria","author":"Nikolaeva D","year":"2015","unstructured":"Nikolaeva D, Draper SJ, Biswas S. Toward the development of effective transmission-blocking vaccines for malaria. Expert Review of Vaccines. Volume 14. Expert Reviews Ltd.; 2015. pp. 653\u201380."},{"issue":"8","key":"ref31","doi-asserted-by":"crossref","first-page":"1343","DOI":"10.1128\/CVI.05104-11","article-title":"N-terminal prodomain of Pfs230 synthesized using a cell-free system is sufficient to induce complement-dependent malaria transmission-blocking activity","volume":"18","author":"Tachibana M","year":"2011","unstructured":"Tachibana M, Wu Y, Iriko H, Muratova O, MacDonald NJ, Sattabongkot J, et al. N-terminal prodomain of Pfs230 synthesized using a cell-free system is sufficient to induce complement-dependent malaria transmission-blocking activity. Clin Vaccine Immunol. 2011;18(8):1343\u201350.","journal-title":"Clin Vaccine Immunol"},{"key":"ref32","author":"Singh SK","year":"2019","unstructured":"Singh SK, Thrane S, Chourasia BK, Teelen K, Graumans W, Stoter R et al. Pfs230 and Pfs48\/45 fusion proteins elicit strong transmission-blocking antibody responses against plasmodium falciparum. Front Immunol. 2019;10(JUN)."},{"issue":"7","key":"ref33","doi-asserted-by":"crossref","DOI":"10.1172\/JCI146221","article-title":"Pfs230 yields higher malaria transmission-blocking vaccine activity than Pfs25 in humans but not mice","volume":"131","author":"Healy SA","year":"2021","unstructured":"Healy SA, Anderson C, Swihart BJ, Mwakingwe A, Gabriel EE, Decederfelt H et al. Pfs230 yields higher malaria transmission-blocking vaccine activity than Pfs25 in humans but not mice. J Clin Invest. 2021;131(7).","journal-title":"J Clin Invest"},{"key":"ref34","author":"Salinas ND","year":"2023","unstructured":"Salinas ND, Ma R, Dickey TH, McAleese H, Ouahes T, Long CA et al. A potent and durable malaria transmission-blocking vaccine designed from a single-component 60-copy Pfs230D1 nanoparticle. NPJ Vaccines [Internet]. 2023;8(1):124. Available from: https:\/\/www.nature.com\/articles\/s41541-023-00709-8."},{"key":"ref35","first-page":"880","volume-title":"Antibody Therapy Goes to Insects: Monoclonal Antibodies Can Block Plasmodium Transmission to Mosquitoes","author":"Coelho CH","year":"2020","unstructured":"Coelho CH, Jore MM, Canepa GE, Barillas-Mury C, Bousema T, Duffy PE. Antibody Therapy Goes to Insects: Monoclonal Antibodies Can Block Plasmodium Transmission to Mosquitoes. Trends in Parasitology. Volume 36. Elsevier Ltd; 2020. pp. 880\u20133."},{"key":"ref36","volume-title":"Plasmodium 6-Cysteine Proteins: Functional Diversity, Transmission-Blocking Antibodies and Structural Scaffolds","author":"Lyons FMT","year":"2022","unstructured":"Lyons FMT, Gabriela M, Tham WH, Dietrich MH. Plasmodium 6-Cysteine Proteins: Functional Diversity, Transmission-Blocking Antibodies and Structural Scaffolds. Frontiers in Cellular and Infection Microbiology. Volume 12. Frontiers Media S.A.; 2022."},{"key":"ref37","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.2221118120","author":"Carballar-Lejaraz\u00fa R","year":"2023","unstructured":"Carballar-Lejaraz\u00fa R, Dong Y, Pham TB, Tushar T, Corder RM, Mondal A et al. Dual effector population modification gene-drive strains of the African malaria mosquitoes, Anopheles gambiae and Anopheles coluzzii. Proceedings of the National Academy of Sciences [Internet]. 2023;120(29). https:\/\/pnas.org\/doi\/10.1073\/pnas.2221118120."},{"key":"ref38","author":"Dong Y","year":"2020","unstructured":"Dong Y, Sim\u00f5es ML, Dimopoulos G. Versatile transgenic multistage effector-gene combinations for Plasmodium falciparum suppression in Anopheles [Internet]. 2020. Available from: https:\/\/www.science.org."},{"issue":"3","key":"ref39","doi-asserted-by":"crossref","first-page":"82","DOI":"10.1101\/pdb.prot100297","article-title":"Standard Immunization of Mice, Rats, and Hamsters","volume":"2020","author":"Greenfield EA","year":"2020","unstructured":"Greenfield EA. Standard Immunization of Mice, Rats, and Hamsters. Cold Spring Harb Protoc. 2020;2020(3):82\u20134.","journal-title":"Cold Spring Harb Protoc"},{"issue":"12","key":"ref40","doi-asserted-by":"crossref","DOI":"10.1371\/journal.ppat.1002458","article-title":"Engineered anopheles immunity to plasmodium infection","volume":"7","author":"Dong Y","year":"2011","unstructured":"Dong Y, Das S, Cirimotich C, Souza-Neto JA, McLean KJ, Dimopoulos G. Engineered anopheles immunity to plasmodium infection. PLoS Pathog. 2011;7(12).","journal-title":"PLoS Pathog"},{"issue":"3","key":"ref41","doi-asserted-by":"crossref","first-page":"1312","DOI":"10.1038\/nprot.2006.221","article-title":"High efficiency germ-line transformation of mosquitoes","volume":"1","author":"Lobo NF","year":"2006","unstructured":"Lobo NF, Clayton JR, Fraser MJ, Kafatos FC, Collins FH. High efficiency germ-line transformation of mosquitoes. Nat Protoc. 2006;1(3):1312\u20137.","journal-title":"Nat Protoc"}],"container-title":[],"original-title":[],"link":[{"URL":"https:\/\/www.researchsquare.com\/article\/rs-3757253\/v1","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.researchsquare.com\/article\/rs-3757253\/v1.html","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,1,25]],"date-time":"2024-01-25T18:46:20Z","timestamp":1706208380000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.researchsquare.com\/article\/rs-3757253\/v1"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,12,19]]},"references-count":41,"URL":"https:\/\/doi.org\/10.21203\/rs.3.rs-3757253\/v1","relation":{},"subject":[],"published":{"date-parts":[[2023,12,19]]},"subtype":"preprint"}}