Influência de diferentes espectros luminosos no desenvolvimento e na produção do fragmento de anticorpo recombinante scFvBaP1 em Nicotiana benthamiana

Abstract

Plant-based molecular farming has become established as a promising platform for obtaining biopharmaceuticals, including recombinant antibodies for the treatment of snakebite envenoming. Optimizing cultivation conditions, such as light spectral quality, represents a strategy to increase yield without the need for further genetic transformation. This study aimed to evaluate the effect of different light spectra – white, Deep Red, and mixed Green Power – on plant development and the production of the recombinant antibody fragment scFv-anti-BaP1, specific against metalloproteinases from Bothrops spp. venom, in homozygous transgenic lines of Nicotiana benthamiana. Initially, five T2 lines were subjected to segregation testing on kanamycin-containing medium, confirming homozygosity in four of the five lines. Plants from these lines were grown under the three spectra for 28 days, with harvests at 14 and 28 days. Morphological parameters, relative transgene expression by RT-qPCR, and recombinant protein accumulation by Western blot followed by densitometry were assessed. Morphological results showed that red light induced significant etiolation at 14 days, remaining superior at 28 days, while white and Green Power lights did not differ from each other. Gene expression revealed effects dependent on the reference gene and time point. The recombinant protein was detected in all treatments and time periods. Although Western blot densitometry did not reveal statistically significant differences among treatments, Dot blot analyses identified higher protein accumulation under the Green Power treatment at both sampling times, differing significantly from the White and Deep Red treatments. These findings indicate that homozygous N. benthamiana lines expressing scFv-anti-BaP1 maintain stable recombinant antibody production under different light spectra while exhibiting distinct responses in protein accumulation. Among the evaluated lighting conditions, the Philips Green Power LED spectrum was associated with the highest levels of recombinant protein accumulation.