Expression of an scFv antibody fragment in Nicotiana benthamiana and in vitro assessment of its neutralizing potential against the snake venom metalloproteinase BaP1 from Bothrops asper

Abstract

Expression of an scFv antibody fragment in Nicotiana benthamiana and in vitro assessment of its neutralizing potential against the snake venom metalloproteinase BaP1 from Bothrops asper

Evaluation of led light on transgenic Nicotiana benthamiana production of a recombinant antibody fragment scFvanti-BAP1.

“…The transgenic plant lines do not show statistical signi cance difference between them or compared to the control plant (WT of N. benthamiana) in terms of nuclear content, although the P2 lineage showed slower growth after 4 weeks, producing less biomass. Thus, P2, P3, and P4 carrying T-DNA scFv-antiBAP1 as T0 plants (Gomes et al, 2019) do not vary in ploidy level as demonstrated by ow cytometry analysis (Fig. 1).…”

“…The total soluble protein extract was puri ed to obtain the scFv-anti-BAP1 by a nity chromatography through the HisTrap nickel column (1 mL). According to the manufacturer's instructions, puri cation was carried out using chromatographic buffer (NaCl and NaHPO 4 ) plus 1M imidazole for elution, as described in Gomes et al (2019). Each experiment was repeated three times.…”

Expression of Single Chain Variable Fragment (scFv) Molecules in Plants: A Comprehensive Update

“…Well-characterized in vitro propagation, transformation, and maintenance methods of these plants are the factors behind their selection. The protein was expressed in whole plant tissue [66,68,69], leaves [70][71][72][73][74][75], flower [76], seeds [77][78][79][80][81], hairy root culture [82] or tobacco BY-2 (tobacco cultivar Bright Yellow) cells [8,83,84] (Fig. 4).…”

“…The fusion product was accumulated in the oil bodies to a range of nearly 0.9% of total seed protein, showed a similar expression profile as that of oleosin [108]. Gomes and co-workers used a SUMO (Small Ubiquitin-like Modifier) fusion protein with a scFv against the metalloproteinase BaP1 (Bothrops asper P1) increased the protein expression in N. benthamiana [84]. The advantages of targeting the protein expression to seeds along with the technical details on expression strategies and the issues to be considered when the product is having a pharmaceutical application have been explained well in one of the pioneering reviews on the seed-specific expression of recombinant proteins [118].…”

Revolutionizing snakebite care with novel antivenoms: Breakthroughs and barriers

“…The pluribody-based formulation, derived from camels, exhibited the ability to neutralize toxin activities and provided effective protection to mice against lethal doses of venom [ 64 ]. Additionally, the study advocates for the utilization of plant cells to produce scFvBaP1, emphasizing its potential as a biotechnological alternative to horse immunization protocols for human therapy against snakebites [ 4 , 65 ].…”