Isolation and light chain shuffling of a Plasmodium falciparum AMA1-specific human monoclonal antibody with growth inhibitory activity

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dc.contributor.authorSeidel-Greven, Melanie
dc.contributor.authorAddai-Mensah, Otchere
dc.contributor.authorSpiegel, Holger
dc.contributor.authorChiegoua Dipah, Gwladys N
dc.contributor.authorSchmitz, Stefan
dc.contributor.authorBreuer, Gudrun
dc.contributor.authorFrempong, Margaret
dc.contributor.authorReimann, Andreas
dc.contributor.authorKlockenbring, Torsten
dc.contributor.authorFischer, Rainer
dc.contributor.authorBarth, Stefan
dc.contributor.authorFendel, Rolf
dc.date.accessioned2021-10-11T09:32:38Z
dc.date.available2021-10-11T09:32:38Z
dc.date.issued2021-01-11
dc.date.updated2021-01-17T04:08:38Z
dc.description.abstractAbstract Background Plasmodium falciparum, the parasite causing malaria, affects populations in many endemic countries threatening mainly individuals with low malaria immunity, especially children. Despite the approval of the first malaria vaccine Mosquirix™ and very promising data using cryopreserved P. falciparum sporozoites (PfSPZ), further research is needed to elucidate the mechanisms of humoral immunity for the development of next-generation vaccines and alternative malaria therapies including antibody therapy. A high prevalence of antibodies against AMA1 in immune individuals has made this antigen one of the major blood-stage vaccine candidates. Material and methods Using antibody phage display, an AMA1-specific growth inhibitory human monoclonal antibody from a malaria-immune Fab library using a set of three AMA1 diversity covering variants (DiCo 1–3), which represents a wide range of AMA1 antigen sequences, was selected. The functionality of the selected clone was tested in vitro using a growth inhibition assay with P. falciparum strain 3D7. To potentially improve affinity and functional activity of the isolated antibody, a phage display mediated light chain shuffling was employed. The parental light chain was replaced with a light chain repertoire derived from the same population of human V genes, these selected antibodies were tested in binding tests and in functionality assays. Results The selected parental antibody achieved a 50% effective concentration (EC50) of 1.25 mg/mL. The subsequent light chain shuffling led to the generation of four derivatives of the parental clone with higher expression levels, similar or increased affinity and improved EC50 against 3D7 of 0.29 mg/mL. Pairwise epitope mapping gave evidence for binding to AMA1 domain II without competing with RON2. Conclusion We have thus shown that a compact immune human phage display library is sufficient for the isolation of potent inhibitory monoclonal antibodies and that minor sequence mutations dramatically increase expression levels in Nicotiana benthamiana. Interestingly, the antibody blocks parasite inhibition independently of binding to RON2, thus having a yet undescribed mode of action.
dc.identifier.ris TY - AU - Seidel-Greven, Melanie AU - Addai-Mensah, Otchere AU - Spiegel, Holger AU - Chiegoua Dipah, Gwladys N AU - Schmitz, Stefan AU - Breuer, Gudrun AU - Frempong, Margaret AU - Reimann, Andreas AU - Klockenbring, Torsten AU - Fischer, Rainer AU - Barth, Stefan AU - Fendel, Rolf AB - Abstract Background Plasmodium falciparum, the parasite causing malaria, affects populations in many endemic countries threatening mainly individuals with low malaria immunity, especially children. Despite the approval of the first malaria vaccine Mosquirix™ and very promising data using cryopreserved P. falciparum sporozoites (PfSPZ), further research is needed to elucidate the mechanisms of humoral immunity for the development of next-generation vaccines and alternative malaria therapies including antibody therapy. A high prevalence of antibodies against AMA1 in immune individuals has made this antigen one of the major blood-stage vaccine candidates. Material and methods Using antibody phage display, an AMA1-specific growth inhibitory human monoclonal antibody from a malaria-immune Fab library using a set of three AMA1 diversity covering variants (DiCo 1–3), which represents a wide range of AMA1 antigen sequences, was selected. The functionality of the selected clone was tested in vitro using a growth inhibition assay with P. falciparum strain 3D7. To potentially improve affinity and functional activity of the isolated antibody, a phage display mediated light chain shuffling was employed. The parental light chain was replaced with a light chain repertoire derived from the same population of human V genes, these selected antibodies were tested in binding tests and in functionality assays. Results The selected parental antibody achieved a 50% effective concentration (EC50) of 1.25 mg/mL. The subsequent light chain shuffling led to the generation of four derivatives of the parental clone with higher expression levels, similar or increased affinity and improved EC50 against 3D7 of 0.29 mg/mL. Pairwise epitope mapping gave evidence for binding to AMA1 domain II without competing with RON2. Conclusion We have thus shown that a compact immune human phage display library is sufficient for the isolation of potent inhibitory monoclonal antibodies and that minor sequence mutations dramatically increase expression levels in Nicotiana benthamiana. Interestingly, the antibody blocks parasite inhibition independently of binding to RON2, thus having a yet undescribed mode of action. DA - 2021-01-11 DB - OpenUCT DP - University of Cape Town IS - Article number: 37 J1 - Malaria Journal KW - Malaria KW - Plasmodium falciparum KW - Apical membrane antigen 1 KW - Human monoclonal antibodies KW - Parasite growth inhibition KW - Phage display KW - Chain shuffling KW - Plant expression LK - https://open.uct.ac.za PY - 2021 T1 - Isolation and light chain shuffling of a Plasmodium falciparum AMA1-specific human monoclonal antibody with growth inhibitory activity TI - Isolation and light chain shuffling of a Plasmodium falciparum AMA1-specific human monoclonal antibody with growth inhibitory activity UR - http://hdl.handle.net/11427/35151 ER - en_ZA
dc.identifier.urihttps://doi.org/10.1186/s12936-020-03548-3
dc.identifier.urihttp://hdl.handle.net/11427/35151
dc.language.rfc3066en
dc.publisher.departmentInstitute of Infectious Disease & Molecular Medicine
dc.publisher.facultyFaculty of Health Sciences
dc.rights.holderThe Author(s)
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/
dc.sourceMalaria Journal
dc.source.journalissueArticle number: 37
dc.source.journalvolume20
dc.source.urihttps://malariajournal.biomedcentral.com/
dc.subjectMalaria
dc.subjectPlasmodium falciparum
dc.subjectApical membrane antigen 1
dc.subjectHuman monoclonal antibodies
dc.subjectParasite growth inhibition
dc.subjectPhage display
dc.subjectChain shuffling
dc.subjectPlant expression
dc.titleIsolation and light chain shuffling of a Plasmodium falciparum AMA1-specific human monoclonal antibody with growth inhibitory activity
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