Plant Recycling for Molecular Biofarming to Produce Recombinant Anti-Cancer mAb

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.

Plant molecular farming: production of metallic nanoparticles and therapeutic proteins using green factories

“…The approach is completely flexible since the specific part of it could be easily re-designed for various targets. 194 Kim et al 195 targeting approaches for recombinant protein accumulation. [196][197][198] Importantly, in one study, in order to find the best extraction method for recombinant monoclonal antibodies from tobacco plants, various factors have been evaluated including pH, temperature, mechanical disruption method, buffer composition, etc.…”

“…Selection of the best plant species for steady alteration and management of environmental parameters which influence plant fitness conditions are very critical. 114,195,[207][208][209] In one study, Lim et al 210 demonstrated that colorectal cancer vaccine protein levels in stems and leaves reaped after flower fertilization were smaller than the plant material gathered before the blossoming period. As a result, the highest manifestation level of a colorectal cancer vaccine protein was reported in the 12 weeks after the in vitro plant seedlings were transplanted.…”

“…were articulated in plants, and accordingly, non-Hodgkin's lymphoma, colorectal cancer, and cervical cancer were targeted. 114,195,[207][208][209] One of the important candidates for anticancer vaccine production is E7 oncoprotein from Human Papilloma Virus (HPV). Massa et al reported HPV16 E7 coding sequence (wild type or mutagenized sequence, E7GGG) as fusions to β-1,3-1,4-glucanase (LicKM) of Clostridium thermocellum and produced in Nicotiana benthamiana plants using a transient expression system.…”

“…reported patterns for the glycosylation and expression of monoclonal antibody CO17-1A (as colorectal anticancer agent) identifying the tumor-associated antigen GA733-2, expressed in human colorectal carcinoma cells in the stem and leaf tissues of primary (0 cycle), secondary (1 cycle), and tertiary (2 cycle) growths of seedlings achieved from the stem cut of T2 plants 195. Consequently, in the 1 and 2 cycle growths, the stages for floral organ creation (35 days) was briefer than that (100 days) for the 0 cycle growth.…”

Endoplasmic Reticulum Retention Motif Fused to Recombinant Anti-cancer Monoclonal Antibody (mAb) CO17-1A Affects mAb Expression and Plant Stress Response

“…We obtained homozygous seeds from transgenic Arabidopsis thaliana plants expressing anti-CRC mAb P s (mAb CO and mAb CO tagged with KDEL (mAb COK)) to recognize CRC-associated antigen GA733 (EpCAM), which is highly expressed in CRC cells [18, 19]. To confirm the effects of ER retention motif tagging on transgenic Arabidopsis , the KDEL motif was fused to the C-terminus of the original HC.…”