Cellular engineering of plant cells for improved therapeutic protein production

Abstract

In vitro cultured plant cells, in particular the tobacco BY-2 cell, have demonstrated their potential to provide a promising bioproduction platform for therapeutic proteins by integrating the merits of whole-plant cultivation systems with those of microbial and mammalian cell cultures. Over the past three decades, substantial progress has been made in improving the plant cell culture system, resulting in a few commercial success cases, such as taliglucerase alfa (Elelyso ® ), the first FDA-approved recombinant pharmaceutical protein derived from plant cells. However, compared to the major expression hosts (bacteria, yeast, and mammalian cells), plant cells are still largely underutilized, mainly due to low productivity and non-human glycosylation. Modern molecular biology tools, in particular RNAi and the latest genome editing technology CRISPR/Cas9, have been used to modulate the genome of plant cells to create new cell lines that exhibit desired “traits” for producing therapeutic proteins. This review highlights the recent advances in cellular engineering of plant cells towards improved recombinant protein production, including creating cell lines with deficient protease levels or humanized glycosylation, and considers potential development in the future.

Engineering Considerations to Produce Bioactive Compounds from Plant Cell Suspension Culture in Bioreactors

“…Plant cell culture (PCC) is nowadays recognized as a promising, renewable, sustainable, and environmentally friendly alternative to obtain PDSM out of wild plants. PCC accounts for the virtues of whole-plant cultivation systems and offers significant advantages, such as controlled manufacture due to standardized environmental conditions, i.e., it is not seasonal dependent, makes use of low amounts of water, and pesticides and herbicides are not required, achieving better quality in the desired product [42,43]. The establishment of different types of plant cell cultures are aimed for regenerating organs or somatic embryos for plant propagation, virus elimination, genetic manipulation, germplasm storage, and for in vitro production of PDSM [44,45].…”

Assessment of the Level of Accumulation of the dIFN Protein Integrated by the Knock-In Method into the Region of the Histone H3.3 Gene of Arabidopsis thaliana

“…Moreover, some recombinant proteins, such as immunotoxins or regulatory proteins, are toxic to mammalian cells. Production of biopharmaceuticals in plant expression systems is becoming a promising alternative to existing platforms based on mammalian and bacterial cells [2,5,6]. The cultivation of plant cells under controlled conditions of bioreactors ensures the production of high-quality protein according to GMP standards.…”

“…Plant cell cultures as a system for the production of recombinant proteins, however, have several drawbacks, the main one of which is associated with a low yield of recombinant protein, which rarely exceeds 100 µg/kg of fresh weight, despite significant efforts to optimize protein expression and stability [6]. In whole plants, the problem of low yield of the target protein can be overcome by transient expression.…”

Strategies for Glycoengineering Therapeutic Proteins

“…Therapeutic protein production in plants has several advantages, including that the infrastructure for large scale production of crops is in place and in theory requires only water, sunlight, and cheap fertilizers ( Burnett & Burnett 2020 ; Karki et al, 2021 ). In practice, current plant-based manufacturing uses cell-based methodology rather than field-grown crops but retains advantages over mammalian cell culture.…”