Abstract
The expression and glycosylation patterns of anti-colorectal cancer therapeutic monoclonal antibody (mAb) CO17-1A recognizing the tumor-associated antigen GA733-2, expressed in human colorectal carcinoma cells, were observed in the leaf and stem tissues of primary (0 cycle), secondary (1 cycle), and tertiary (2 cycle) growths of seedlings obtained from the stem cut of T2 plants. The bottom portion of the stem of T2 seedlings was cut to induce the 1 cycle shoot growth, which was again cut to induce the 2 cycle shoot growth. In the 1 and 2 cycle growths, the periods for floral organ formation (35 days) was shorter than that (100 days) for the 0 cycle growth. The genes of heavy and light chains of mAb CO17-1A existed at the top, middle, and basal portions of the leaves and stem obtained from the 0, 1, and 2 cycle plants. The protein levels in the leaves and stem tissues from the 1 and 2 cycles were similar to those in the tissues from the 0 cycle. The glycosylation level and pattern in the leaf and stem did not alter dramatically over the different cycles. Surface plasmon resonance (SPR) confirmed that mAbs CO17-1A obtained from leaf and stem tissues of the 0, 1, and 2 cycles had similar binding affinity for the GA733-2 antigen. These data suggest that the shoot growth by bottom stem cutting is applicable to speed up the growth of plant biomass expressing anti-colorectal cancer mAb without variation of expression, glycosylation, and functionality.
Highlights
Plants are well recognized as alternative hosts for production of highly valuable recombinant proteins, such as antibodies, vaccines, human blood products, hormones, and growth regulators (Fernandez-San Millan et al, 2003; Rigano and Walmsley, 2005; Schillberg et al, 2013)
We demonstrate that fresh stem and leaves regrow from axillary buds after cutting of the stem
Fully grown Nicotiana tabacum plants started to form the floral organs at 12 weeks after sowing (Lim et al, 2015), which is much longer than N. benthamiana (7 weeks; Conley et al, 2011)
Summary
Plants are well recognized as alternative hosts for production of highly valuable recombinant proteins, such as antibodies, vaccines, human blood products, hormones, and growth regulators (Fernandez-San Millan et al, 2003; Rigano and Walmsley, 2005; Schillberg et al, 2013). They offer mass production and safety advantages, with ease of seed storage, compared to the microbial and animal cell-based systems (Twyman et al, 2003; Fischer et al, 2013; Shanmugaraj and Ramalingam, 2014). A plant-based recombinant protein production system might have drawbacks, such as relatively long cultivation period for obtaining full biomass from the seeds, especially when there is
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