All ETDs from UAB

Advisory Committee Chair

Renata Jaskula-Sztul

Advisory Committee Members

Xiaoguang Liu

Gangjian Qin

Lufang Zhou

Yang Zhou

Document Type

Dissertation

Date of Award

2022

Degree Name by School

Doctor of Philosophy (PhD) School of Engineering

Abstract

Malignant cancer is one of the greatest threats to human health globally. Conventional (non-targeted) treatments such as chemo-drugs and radiation therapy remain the major choices for cancer patients while always accompanied by severe systemic toxicity/side effects. Monoclonal antibody (mAb) is one of the most successful biomolecules discovered and well investigated as a targeted therapy to treat human disease. This dissertation research was aimed to develop monoclonal antibody-based targeted drugs for cancer treatment, which include antibody-drug conjugates (ADCs), liposomal drug and exosome-based drug delivery. Novel targets were discovered, screened and confirmed by proteomics, western blotting and flow cytometry analysis. Cancer cells specific mAbs (anti-SSTR2, CD47, CD276) were developed using hybridoma technique and top clones were selected and amplified. Binding affinity was tested by ELISA-based Kd quantitation assay. Internalization function was confirmed by live-cell confocal microscopy on cancer cell lines. Antibodies and toxic drugs were conjugated via chemical linkers (SMCC, MC-Val-Cit-PAB and re-bridging, etc.) to form ADCs. SDS-PAGE, hydrophobic interaction chromatography (HIC) and in vitro cytotoxicity analysis were conducted for ADC characterization. Patient-derived xenograft (PDX) and cell-derived xenograft (CDX) animal models were established for anti-cancer efficacy evaluation. Liposome and exosome were also engineered to achieve mAb-mediated targeted drug delivery. Neutral charged liposome was synthesized by rehydrating dry iv lipid film combined with sonication. While exosome was naturally secreted by live cells and harvested for drug packing. 1,2-Distearoyl-sn-glycero-3-phosphoethanolamine-Poly (ethylene glycol)/DSPE-PEG was utilized to modify the lipid membrane of liposome and exosome to enhance their retention time in vivo and 1,2-Distearoyl-sn-glycero-3-phosphoethanolamine-NHS ester/DSPE-NHS linked tumor-specific mAb to the drug-loaded vesicles. Transmission electron microscopy (TEM) and nanoparticle tracking assay (NTA) were performed for particle characterization. The drug delivery attributes such as lipid particle-cell membrane fusion, tumor-targeting function were confirmed by live-cell confocal microscopy imaging and in vivo imaging system (IVIS). In vivo efficacy of both liposomal drug and exosome-drug were investigated on xenograft animal models. Inspired by promising results, biomanufacturing process was developed which could support large-scale/preclinical animal studies. This dissertation research study expanded the application of monoclonal antibody and exploited new directions for novel targeted therapies development.

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