Author ORCID Identifier
Document Type
Thesis
Date of Award
Spring 4-29-2025
Keywords
antibody-drug conjugate, effector functions, conjugation, glycosylation, Surface Plasmon Resonance
Degree Name
Biochemistry (BS)
Department
BIOCHEMISTRY
First Advisor
L. Nathan Tumey
Abstract
Increasing research has been conducted on antibody drug conjugates (ADCs) for their oncology applications and potential as treatments for immunological diseases. ADCs usually covalently bind a cytotoxic drug (payload) via a linker to a monoclonal antibody, which targets specific antigens expressed on tumors, delivering the drug directly to cancer cells while minimizing off-target effects. In addition to binding antigens on tumor cells, antibodies also interact with immune cells to activate “effector functions”. This activation is largely driven by the binding of Fcγ-receptors (FcγR) to the Fc region of IgG, which connects the humoral immune responses and cell-mediated immune responses. Antibody characteristics, such as glycosylation and conjugation, affect how well they bind to FcγR.
In this paper, both heterogeneous endogenous cysteine ADCs and site-specific ADCs are investigated. Surface plasmon resonance (SPR) was performed to evaluate the binding of naked antibodies, deglycosylated (DG) naked antibodies, glycosylated ADCs, and deglycosylated (DG) ADCs to four different FcγR isotypes. This experiment was performed with HER2 ADCs. Additionally, the dissociation constant (KD) for HER2 samples was also determined to assess the binding affinity of the antibody and ADCs to each FcγR. When the small molecule N-(2-Hydroxyethyl)maleimide was conjugated to a HER2 antibody, the amount of binding to all four FcγRs decreased compared to that of the naked antibody. Furthermore, all the deglycosylated samples exhibited a lower amount of FcγR binding than their glycosylated counterparts.
Effector function assays were performed to evaluate antibody-dependent cellular cytotoxicity (ADCC) and antibody-dependent cellular phagocytosis (ADCP). Consistent with the SPR experiment, deglycosylated HER2 and TROP2 samples exhibited reduced ADCC and ADCP activity compared to their glycosylated counterparts. Most deglycosylated samples showed little induction of ADCC or ADCP.
Recommended Citation
Wu, Lina, "Assessing the Effects of Antibody Glycosylation and Conjugation on Fc-mediated Effector Functions" (2025). Undergraduate Honors Theses. 55.
https://orb.binghamton.edu/undergrad_honors_theses/55
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