Preprint: Characterisation of a Highly Potent and Near Pan-Neutralising Anti-HIV Monoclonal Antibody Expressed in Tobacco Plants

Catherine Margaret Moore, Melanie Grandits, Clemens Grünwald-Gruber, Friedrich Altmann, Maria Kotouckova, Audrey Y.H. Teh and Julian K.C. Ma 2020. Preprint: Characterisation of a Highly Potent and Near Pan-Neutralising Anti-HIV Monoclonal Antibody Expressed in Tobacco Plants. Research Square. https://doi.org/10.21203/rs.3.rs-104508/v1

TitlePreprint: Characterisation of a Highly Potent and Near Pan-Neutralising Anti-HIV Monoclonal Antibody Expressed in Tobacco Plants
AuthorsCatherine Margaret Moore, Melanie Grandits, Clemens Grünwald-Gruber, Friedrich Altmann, Maria Kotouckova, Audrey Y.H. Teh and Julian K.C. Ma
Description

Background HIV remains one of the most important health issues worldwide, with almost 40 million people living with HIV. Although patients develop antibodies against the virus, its high mutation rate allows evasion of immune responses. Some patients, however, produce antibodies that are able to bind to, and neutralise different strains of HIV. One such ‘broadly neutralising’ antibody is ‘N6’. Identified in 2016, N6 can neutralise 98% of HIV-1 isolates with a median IC50 of 0.066 µg/mL. This neutralisation breadth makes N6 a very promising therapeutic candidate.

Results N6 was expressed in a glycoengineered line of N. benthamiana plants (pN6) and compared to the mammalian cell-expressed equivalent (mN6). Expression at 49mg/kg (fresh leaf tissue) was achieved in plants, although extraction and purification are more challenging than for most plant-expressed antibodies. N-glycoanalysis demonstrated the absence of xylosylation and a reduction in α(1,3)-fucosylation that are typically found in plant glycoproteins. The N6 light chain contains a potential N-glycosylation site, which was modified and displayed more α(1,3)-fucose than the heavy chain. The binding kinetics of pN6 and mN6, measured by surface plasmon resonance, were similar for HIV gp120. pN6 had a 10-fold higher affinity for FcγRIIIa, which was reflected in an antibody-dependent cellular cytotoxicity assay, where pN6 induced a more potent response from effector cells than that of mN6. pN6 demonstrated the same potency and breadth of neutralisation as mN6, against a panel of HIV strains.

Conclusions The successful expression of N6 in tobacco supports the prospect of developing a low-cost, low-tech production platform for a monoclonal antibody cocktail to control HIV in low-to middle income countries.

Year2020
Output mediaPreprint, made available on Research Square
PublisherResearch Square
Publication dates
Published online17 Nov 2020
Digital Object Identifier (DOI)https://doi.org/10.21203/rs.3.rs-104508/v1
Web address (URL)https://doi.org/10.21203/rs.3.rs-104508/v1

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