Plant-based production of highly potent anti-HIV antibodies with engineered posttranslational modifications

Singh, Advaita AcaryaPooe, OKwezi, LusisizweLotter-Stark, TStoychev, Stoyan HAlexandra, Kabamba BGerber, Isak BBhiman, JNVorster, JPauly, MZeitlin, LWhaley, KMach, LSteinkellner, HMorris, LTsekoa, Tsepo LChikwamba, Rachel K2021-06-232021-06-232020-04Singh, A.A., Pooe, O., Kwezi, L., Lotter-Stark, T., Stoychev, S.H., Alexandra, K.B., Gerber, I.B. & Bhiman, J. et al. 2020. Plant-based production of highly potent anti-HIV antibodies with engineered posttranslational modifications. Scientific Reports, 10(2601). http://hdl.handle.net/10204/120252045-2322DOI.org/10.1038/s41598-020-63052-1http://hdl.handle.net/10204/12025Broadly neutralising antibodies (bNAbs) against human immunodeficiency virus type 1 (HIV-1), such as CAP256-VRC26 are being developed for HIV prevention and treatment. These Abs carry a unique but crucial post-translational modification (PTM), namely O-sulfated tyrosine in the heavy chain complementarity determining region (CDR) H3 loop. Several studies have demonstrated that plants are suitable hosts for the generation of highly active anti-HIV-1 antibodies with the potential to engineer PTMs. Here we report the expression and characterisation of CAP256-VRC26 bNAbs with posttranslational modifications (PTM). Two variants, CAP256-VRC26 (08 and 09) were expressed in glycoengineered Nicotiana benthamiana plants. By in planta co-expression of tyrosyl protein sulfotransferase 1, we installed O-sulfated tyrosine in CDR H3 of both bNAbs. These exhibited similar structural folding to the mammalian cell produced bNAbs, but non-sulfated versions showed loss of neutralisation breadth and potency. In contrast, tyrosine sulfated versions displayed equivalent neutralising activity to mammalian produced antibodies retaining exceptional potency against some subtype C viruses. Together, the data demonstrate the enormous potential of plant-based systems for multiple posttranslational engineering and production of fully active bNAbs for application in passive immunisation or as an alternative for current HIV/AIDS antiretroviral therapy regimens.FulltextenAnti-HIV antibodiesBroadly neutralising antibodiesbNAbsPlant-based production of highly potent anti-HIV antibodies with engineered posttranslational modificationsArticleSingh, A. A., Pooe, O., Kwezi, L., Lotter-Stark, T., Stoychev, S. H., Alexandra, K. B., ... Chikwamba, R. K. (2020). Plant-based production of highly potent anti-HIV antibodies with engineered posttranslational modifications. Scientific Reports, 10(2601), http://hdl.handle.net/10204/12025Singh, Advaita Acarya, O Pooe, Lusisizwe Kwezi, T Lotter-Stark, Stoyan H Stoychev, Kabamba B Alexandra, Isak B Gerber, et al "Plant-based production of highly potent anti-HIV antibodies with engineered posttranslational modifications." Scientific Reports, 10(2601) (2020) http://hdl.handle.net/10204/12025Singh AA, Pooe O, Kwezi L, Lotter-Stark T, Stoychev SH, Alexandra KB, et al. Plant-based production of highly potent anti-HIV antibodies with engineered posttranslational modifications. Scientific Reports, 10(2601). 2020; http://hdl.handle.net/10204/12025.TY - Article AU - Singh, Advaita Acarya AU - Pooe, O AU - Kwezi, Lusisizwe AU - Lotter-Stark, T AU - Stoychev, Stoyan H AU - Alexandra, Kabamba B AU - Gerber, Isak B AU - Bhiman, JN AU - Vorster, J AU - Pauly, M AU - Zeitlin, L AU - Whaley, K AU - Mach, L AU - Steinkellner, H AU - Morris, L AU - Tsekoa, Tsepo L AU - Chikwamba, Rachel K AB - Broadly neutralising antibodies (bNAbs) against human immunodeficiency virus type 1 (HIV-1), such as CAP256-VRC26 are being developed for HIV prevention and treatment. These Abs carry a unique but crucial post-translational modification (PTM), namely O-sulfated tyrosine in the heavy chain complementarity determining region (CDR) H3 loop. Several studies have demonstrated that plants are suitable hosts for the generation of highly active anti-HIV-1 antibodies with the potential to engineer PTMs. Here we report the expression and characterisation of CAP256-VRC26 bNAbs with posttranslational modifications (PTM). Two variants, CAP256-VRC26 (08 and 09) were expressed in glycoengineered Nicotiana benthamiana plants. By in planta co-expression of tyrosyl protein sulfotransferase 1, we installed O-sulfated tyrosine in CDR H3 of both bNAbs. These exhibited similar structural folding to the mammalian cell produced bNAbs, but non-sulfated versions showed loss of neutralisation breadth and potency. In contrast, tyrosine sulfated versions displayed equivalent neutralising activity to mammalian produced antibodies retaining exceptional potency against some subtype C viruses. Together, the data demonstrate the enormous potential of plant-based systems for multiple posttranslational engineering and production of fully active bNAbs for application in passive immunisation or as an alternative for current HIV/AIDS antiretroviral therapy regimens. DA - 2020-04 DB - ResearchSpace DP - CSIR J1 - Scientific Reports, 10(2601) KW - Anti-HIV antibodies KW - Broadly neutralising antibodies KW - bNAbs LK - https://researchspace.csir.co.za PY - 2020 SM - 2045-2322 T1 - Plant-based production of highly potent anti-HIV antibodies with engineered posttranslational modifications TI - Plant-based production of highly potent anti-HIV antibodies with engineered posttranslational modifications UR - http://hdl.handle.net/10204/12025 ER -23778