[CCoE Notice] Cullen College Dissertation Defense Announcement - Atul Goyal

[Knudsen, Rachel W]

[Dissertation Defense Announcement at the Cullen College of Engineering]
Detection of Biomolecules using Novel Approaches

Atul Goyal
August 2, 2022;  10:30 AM (CST)

Room: Chemical Engineering Conference Room
Zoom Meeting ID:
 https://urldefense.com/v3/__https://us02web.zoom.us/j/85412637467pwd=Q1dHOFBnY1RXYXhHUmw2dmkvaEZSUT09__;!!LkSTlj0I!DoRpVD8yVdaGOa2jxEXfz8FHBVcAiFyDEOfFzkcgUTW7iR6aFMLktPuoGL-_8nTfhTFRsCpJif7kaSlLa6h083zk$ <https://urldefense.com/v3/__https:/us02web.zoom.us/j/85412637467pwd=Q1dHOFBnY1RXYXhHUmw2dmkvaEZSUT09__;!!LkSTlj0I!GU4M33VF95bEGI4eSpyeQQyVGrfWNAkF_Oux654niyANqAxE-YWZpH7aV3nuIEphzTU6P11L54KcDmpoPZnWmuOWS24$>


Committee Chair:
Richard Willson, Ph.D.
Committee Members:
Navin Varadarajan, Ph.D. | Gul Zerze, Ph.D. | Jiming Bao, Ph.D. | Paul Ruchhoeft, PhD.
Abstract
In the manufacture of therapeutic monoclonal antibodies (mAbs), the clarified cell culture fluid is typically loaded onto an initial protein A affinity capture column. Imperfect mass transfer and loading to maximum capacity can risk antibody breakthrough and loss of valuable product, but conservative underloading wastes expensive protein A resin. In addition, the effects of column fouling and ligand degradation require the frequent optimization of IgG loading to avoid wastage. Therefore, continuous real-time monitoring of IgG flowthrough is of great interest. We previously developed a fluorescence-based monitoring technology that allows mix-and-read mAb detection in cell culture fluid. Here we report the use of reporters immobilized on CNBr-activated Sepharose 4B resin for continuous detection of IgG in column breakthrough. The column effluent is continuously contacted with immobilized fluorescein-labeled Fc-binding ligands to produce an immediately detectable change in fluorescence intensity. The technology allows rapid and reliable monitoring of IgG in a flowing stream of clarified cell culture fluid emerging from a protein A column, without prior sample preparation. We observed a significant change in fluorescence intensity at 0.5 g/L human IgG, sufficient to detect a 5% breakthrough of a 10 g/L load, within 2 minutes at a flow rate of 0.5 mL/min.
In another study, we investigated the affinity capture of nanoparticles using membrane chromatography. A major cause of the continuous wide spread of COVID-19 is the lack of a fast, cheap, and reliable diagnostic technique. For the most reliable COVID-19 result, the PCR testing still has to be performed by medical specialists at properly-equipped sites, and both patients and health authorities sometimes have to wait for days to get the results, significantly hampering efforts to control and minimize the spread of the virus. This research aimed to develop an ultra-sensitive detection method for the novel coronavirus and other analytes that is inexpensive, portable, and does not require PCR amplification or its associated expensive equipment. We used micro and nanoscale holes through which light can pass and which can be occluded by single nanoparticles as small as 110 nm in diameter. While chemistry for the successful affinity capture of antibody conjugated nanoparticles to effectively deliver to nanoholes for detection was tested, it continues to be developed..
[Engineered For What's Next]

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