Biomedical Engineering and Bioengineering Commons^™

Intrabodies Reveal Critical Steps Involved In Ricin's Interactions With The Ribosome, Timothy Francis Czajka

Legacy Theses & Dissertations (2009 - 2024)

Ricin is a highly lethal protein toxin derived from the seeds of the castor plant, Ricinus communis. It is a Type II ribosome inactivating protein (RIP), meaning it is a heterodimer with one subunit, ricin toxin B (RTB), that mediates cell surface attachment and intracellular trafficking and a second subunit, ricin toxin A (RTA), that irreversibly shuts down protein synthesis in the cytosol. During trafficking, RTA and RTB necessarily separate in the endoplasmic reticulum, wherein RTA unfolds and translocates into the cytosol where it refolds into an enzymatically active conformation. RTA is remarkably fast acting and efficient, with few molecules …

Quantification Of Tumor-Associated Macrophages Following Immunomodulation Therapy In A Murine Allograft Model Of Colorectal Carcinoma, Caroline Spainhour

Biomedical Engineering Undergraduate Honors Theses

Colorectal Carcinoma (CRC) is one of the deadliest cancers in the world, with 150,000 new cases annually in the United States. Traditional treatments include chemotherapy and invasive surgery; however, research has shown that only 25% of patients that undergo traditional treatment have a positive result. Immunotherapy is an emerging form of cancer treatment that utilizes the patients’immune system to fight cancer cells by targeting inflammation, which plays a large role in the proliferation and metastasis of cancer cells.

Tumor-associated macrophages (TAMs) are immune cells that affect the inflammatory microenvironment of tumors. TAMs are M1 in the early stages of tumors, …

Aptamer Based Hybrid-Assay For Early Stage Disease Diagnosis, Riley S. Parsons, Mari A. Ueno

Bioengineering Senior Theses

The objective of the project is to use aptamers, oligonucleotides designed to selectively bind target molecules, to develop a methodology for building an enzyme-linked aptasorbent assay (ELASA). This assay is designed to detect the presence of a target protein nucleolin that is overexpressed on the surface of cancer cells, and it would act as a novel diagnostic method for the disease. The success of our project would also confirm the feasibility of using an ELASA as a diagnostic tool to detect the presence of thrombospondin-1 (TSP-1), a protein known to experience glycosylation changes in human endometrioid ovarian cancer tissue. Our …

Development Of Antibody Detection Methods For Active Product At The Cell Culture Stage, Gregory Walsh, Matthew Schraut, Jon Barricelli, Lada Laenen

Cell Culture Engineering XV

Development of new analytical methods to characterize the manufacturing process became necessary when the site wanted to implement process improvements to increase output from the bioreactor. The bioreactor process is monitored and controlled for dissolved oxygen content, glucose concentration, temperature, etc. and samples of the harvest are analyzed daily using an endpoint assay and a total protein method. Neither of the two methods can detect the total amount of product at the harvest stage; the endpoint assay will only detect active product and the total protein method will not discriminate between the cellular proteins and the active product. We have …

An Innovative Protein Delivery System For Therapeutic Cells, Jean Pascal Lepetit

Cell Culture Engineering XV

Cell therapy has been on the rise in recent years. It consists in the treatment of therapeutic cells to be transferred to human patients, and is applied in multiple pathologies such as hemophilia or muscular dystrophies. Several cell treatment solutions are currently developed, including the use of transcription factors or genome editing technologies. These ex vivo technologies often require gene transfer to express the therapeutic protein in the treated cells. The genes are delivered mainly by transfection or viral transduction, causing regulatory limitations for clinical studies.

A promising strategy is to avoid the use of nucleic acids by direct delivery …

Optimization Of Glycosylation And Charge Distribution Through Culture Parameters And Supplements, Sigma Mostafa, Ventaka Tayi, Shahid Rameez, Nathan Oien, Jaspreet Notey, Brian Baker, Jimmy Smedley, Abhinav Shukla

Cell Culture Engineering XV

Culture parameters are known to have significant impacts on product quality, although these effects are sometimes cell-line dependent and the directionality of the effect has to be determined empirically. We will present data from three case studies where glycosylation or charge distribution was modified to match the reference molecule or to reduce variability. In the first case study, glycan optimization for a biosimilar will be described. Galactose, fucose, and mannose levels were optimized through screening of raw materials and process parameters. A range of media, feed, alternate sugars, metals and other supplements as well as temperature set points were tested. …

Integration Of Continuous Ethanol Precipitation And Flocculation Into Manufacturing Of Antibodies, Alois Jungbauer, Nikolaus Hammerschimidt, Peter Satzer

Integrated Continuous Biomanufacturing II

Precipitation and flocculation are optimal unit operations for continuous capture of proteins from culture supernatant. Precipitation and flocculation can be operated in a real continuous manner. The methods can be combined and after this capture step the precipitate can be stored as a concentrated solution or even as a precipitate. We have developed several precipitation/ flocculation protocols for capture of antibodies from culture supernatant. These include the combinations of CaCl₂ flocculation with either cold ethanol precipitation or PEG precipitation and octanoic acid precipitation with PEG precipitation. The precipitation conditions have been screened in microtiter plates or in case of …

Toward Complete Continuity In Antibody Biomanufacture: Multi-Column Continuous Chromatography For Protein A Capture And Mixed Mode Hydroxyapatite Polishing, Anthony Grabski, Tom Van Oosbree, Beth Hammer, Alla Zilberman, Robert Mierendoff

Integrated Continuous Biomanufacturing II

Monoclonal antibodies (mAbs) are a predominant modality for a broad range of clinical indications including oncology and inflammatory diseases. Increasing manufacturing capacity and decreasing cost per purification campaign are critical factors for making antibody therapies more affordable. Cell culture mAb expression levels have steadily increased over the past ten years with titers of five grams per liter frequently achieved. Higher titers reduce production costs and allow processing of kilogram quantities for clinical trials from single use cell culture vessels. Drawbacks of increased titers include higher levels of aggregates, fragments, variants, and process impurities. These combined titer and impurity burdens further …

Comparing Anti-Vegf Antibodies And Aptamers On Paper Microfluidic-Based Platforms, Katherine Noel Clayton

Master's Theses

The field of microfluidics is expanding into what is known as paper microfluidics. This uses a paper platform rather than materials (i.e. PDMS, PMMA) that are commonly used in microfluidics research. Current devices require an expensive manufacturing process and external sources to power the device. Such devices are not practical in low resource environments. As a consequence, it is the goal of this Thesis to develop a three-dimensional, multiplexed assay chip using nitrocellulose membranes. This device comprises of multiple layers of nitrocellulose membranes with defined fluidic channels. The multiple layers are bound together using double backed tape, and imbedded between …