Neutralizing antibodies against rotavirus produced in transgenically labelled purple tomatoes

Abstract

SummaryEdible fruits are inexpensive biofactories for human health-promoting molecules that can be ingested as crude extracts or partially purified formulations. We show here the production of a model human antibody for passive protection against the enteric pathogen rotavirus in transgenically labelled tomato fruits. Transgenic tomato plants expressing a recombinant human immunoglobulin A (hIgA_2A1) selected against the VP8* peptide of rotavirus SA11 strain were obtained. The amount of hIgA_2A1 protein reached 3.6 ± 0.8% of the total soluble protein in the fruit of the transformed plants. Minimally processed fruit-derived products suitable for oral intake showed anti-VP8* binding activity and strongly inhibited virus infection in an in vitro virus neutralization assay. In order to make tomatoes expressing hIgA_2A1 easily distinguishable from wild-type tomatoes, lines expressing hIgA_2A1 transgenes were sexually crossed with a transgenic tomato line expressing the genes encoding Antirrhinum majus Rosea1 and Delila transcription factors, which confer purple colour to the fruit. Consequently, transgenically labelled purple tomato fruits expressing hIgA_2A1 have been developed. The resulting purple-coloured extracts from these fruits contain high levels of recombinant antirotavirus neutralizing human IgA in combination with increased amounts of health-promoting anthocyanins.

Combinatorial Analysis of Secretory Immunoglobulin A (sIgA) Expression in Plants

“…Even more, certain specific decisions concerning antibody design, as the choice for LC and HC isotypes or for subcellular localization, are rarely made on the basis of an exhaustive experimental analysis. However, it has been repeatedly observed that antibody expression levels diverge dramatically from case to case and that, e.g., the experimental selection of the most stable idiotypes, can bring considerable advantages in terms of yield [15,31]. …”

“…The variable regions of 2A1 antibody were initially selected by phage display against the VP8* peptide of the VP4 protein of the rotavirus SA11 strain capsid [30]. This antibody was previously described in its monomeric format, showing a strong rotavirus neutralization activity [31]. …”

Visual Tracking of Plant Virus Infection and Movement Using a Reporter MYB Transcription Factor That Activates Anthocyanin Biosynthesis

“…Silencing of the transgenically expressed transcription factors Ros1 and Delila was also used as a virus-induced gene silencing reporter in purple tomatoes (Orzaez et al, 2009). In contrast, strategies based on induction of tissue pigmentation are not so common, although recently, anthocyanin accumulation has been proposed as a method for identity preservation in plant biotechnology (Juárez et al, 2011) and as a selection system in plant transformation (Kortstee et al, 2011).…”

Tomato (Solanum lycopersicum L.) in the service of biotechnology

“…Recombinant production of rotavirus antigens in plants has been proposed as an alternative to traditional production platforms. Juárez et al (2012) obtained transgenic tomato plants expressing a recombinant human immunoglobulin A (hIgA_2A1) selected against the VP8* peptide of rotavirus SA11 strain. The amount of hIgA_2A1 protein reached 3.6 ± 0.8 % of the TSP in the fruit of the transformed plants.…”