Graduation Year
2026
Document Type
Open Access Senior Thesis
Degree Name
Bachelor of Arts
Department
Biology
Reader 1
Larry Grill
Reader 2
Aditi Vyas
Terms of Use & License Information
Abstract
Monoclonal antibodies, or mAbs, have recently joined the growing array of biopharmaceuticals used to treat a variety of chronic, autoimmune, and infectious diseases. Antibody therapy is highly customizable, as mAbs can be genetically engineered to target specific disease markers. Traditional systems of producing monoclonal antibodies use mammalian cells, most prominently Chinese Hamster Ovary Cells. While these systems are widely used, they are time-intensive and costly. Because of this, plant-based antibody production systems have risen in prominence, as they provide a more cost-efficient and faster way to generate antibodies. One drawback that has surfaced is that the co-expression of antibody heavy and light chains usually requires two viral vectors, which can result in uneven subunit expression. To address this, the goal of this project was to develop a bidirectional tobacco mosaic virus (TMV)-based plant expression vector for dual gene co-expression. This system uses a single vector and is based on the TMV-derived pTRBO backbone, engineered to contain a bidirectional promoter that can drive dual gene expression simultaneously. Agrobacterium tumefaciens was transformed using the pTRBO bidirectional promoter and potato virus X plasmid, which were grown from stocks. The plasmid was inoculated with kanamycin and extracted, and a restriction enzyme digest was performed using CutSmart, NotI-HF, and AvrII. Five Nicotiana benthamiana plants were infiltrated with the digested samples, along with fluorescent reporters GFP, DsRed, or GFP/DsRed, and incubated for two days. Fluorescence microscopy indicated successful insertion of the promoter and multiple reporter gene combinations into plant leaves. A fluorescence time trial was performed in order to assess the longevity of GFP and DsRed in the plants over six days post-infiltration, indicating the relative expression of recombinant proteins.The results of this trial showed high and consistent protein expression levels at six days post-infiltration. These results indicate that the system is able to successfully express recombinant proteins at a consistent level without substantial decreases in production. This suggests the system can be used to produce monoclonal antibodies for future disease treatments. This research could be impactful especially in the field of women’s health, as cervical cancer death rates remain high, and researchers look for ways to combat the spread of HIV, HPV, and other sexually transmitted infections that negatively impact women worldwide.
Recommended Citation
Schutten-Burgess, Matisse, "Development of a Bidirectional TMV-Based Plant Expression Vector for Dual Gene Co-Expression" (2026). Scripps Senior Theses. 2776.
https://scholarship.claremont.edu/scripps_theses/2776
