Continued expression of plant-made vaccines following long-term cryopreservation of antigen-expressing tobacco cell cultures

Abstract

Production of vaccines in plant cells provides an alternative system that has several advantages when compared to current vaccine production methods. Establishment of stable seed stocks for a continuous supply of a vaccine is a critical part of production systems. Therefore, a vitrification method for cryopreservation was applied to non-transgenic and three different antigen-expressing transgenic Nicotiana tabacum (NT-1) lines. Preculture of the suspension cultures 1 d prior to vitrification was sufficient for cell survival through the cryopreservation process. Inclusion of 0.3 M mannitol in the preculture medium was necessary for maintenance of cell viability. Cultures were also treated with and without heat shock prior to vitrification, and it was found that heat shock was unnecessary for growth recovery post cryopreservation. All cultures survived storage in liquid nitrogen at intervals ranging from 1 h to 1 yr. Antigen expression was measured by enzyme-linked immunosorbent assay for cultures that grew post cryopreservation and those that had never been cryopreserved. Expression levels in cultures derived from cryopreserved material were comparable to cultures that had not been cryopreserved. Transmission electron microscopy showed that the integrity of the cell structure was maintained post cryopreservation.

Recent advances towards development and commercialization of plant cell culture processes for the synthesis of biomolecules

“…, 1991). As a result, seed stocks can be established using cryopreservation to limit production variability among batches (Van Eck and Keen, 2009).…”

Plant-Produced Vaccines: Future Applications in Aquaculture

“…Plant-made vaccines have been widely reported against rabies (Ashraf et al, 2005;Loza-Rubio et al, 2008;Roy et al, 2010;Loza-Rubio et al, 2012), porcine reproductive and respiratory syndrome virus, and porcine post-weaning diarrhea in piglets (Chen and Liu, 2011;Kolotilin et al, 2012). A tobacco-made Newcastle disease vaccine for poultry is the only plant-made vaccine approved by the United States Department of Agriculture so far (Hahn et al, 2007;Li et al, 2007;Yang et al, 2007;Joensuu et al, 2008;Gómez et al, 2009;Van Eck and Keen, 2009;Wu et al, 2009;Ma et al, 2020a). Plant-made veterinary vaccines have also been studied in mink, dogs, and cats (Dalsgaard et al, 1997;Molina et al, 2004Molina et al, , 2005Rybicki, 2010).…”

How can plant genetic engineering contribute to cost-effective fish vaccine development for promoting sustainable aquaculture?

“…Each system has its advantages and limitations and the method of choice is largely depending on what kind of fish vaccines are to be expressed, as briefly described in Table 2. To date, both food and non-food crops (especially tobacco plant) have been used for the development of a number of animal vaccines, such as a poultry vaccine against Newcastle disease (Hahn et al 2007; Yang et al 2007; Li et al 2007; Gómez et al2009; Van Eck and Keen 2009; Wu et al 2009: for reviews see Floss et al 2007 and He et al 2008), rabies (Ashraf et al 2005; Loza-Rubio et al 2008; Roy et al 2010; Loza-Rubio et al 2012), Porcine reproductive and respiratory syndrome virus (PRRSV) and Porcine post-weaning diarrhea in piglets (Chen and Liu 2011; Kolotilin et al 2012). The vaccine against Newcastle disease was the first plant-made animal vaccine receiving regulatory approval from the US Department of Agriculture (USDA) Center for Veterinary Biologics in 2006 (www.thepoultrysite.com/poultrynews/8949/usda-issues-license-for-plant-cell-producednewcastle-disease-vaccine-for-chickens; Joensuu et al 2008).…”