The long term objectives of this project are to understand how changes in protein glycosylation regulate antibody dependent immune responses, and then to utilize that knowledge to develop improved antibody-based therapies for diseases such as cancer.
The specific aims of this project are to: 1. Develop methods to produce homogeneous glycoproteins for the study and optimization of antibody glycosylation. 2. Determine the role of N-linked glycosylation in IgG Fc interactions with Fc receptors. 3. Develop methods to incorporate site-specific chemical modifications onto glycosylated IgG Fcs to facilitate biochemical studies and to modify bioactivity. 4. Determine the role of N-linked glycosylation in regulating antibody dependent cellular immune responses. As part of this research, novel methods will be developed to produce homogeneously glycosylated antibody fragments of the immunoglobulin G (IgG) subclasses, and also to attach small molecule receptor ligands to those antibody fragments site-selectively. The glycosylated and chemically modified antibody fragments thus produced will be utilized in in vitro biochemical and structural studies of antibody interactions with Fc receptors to determine the role of glycosylation and IgG subclass in regulating cellular immune responses. Antibody fragments attached to receptor ligands will be utilized to target cancer cells for antibody dependent immune responses in antibody dependent cell-mediated cytotoxicity (ADCC) assays and complement dependent cytotoxicity (CDC) assays. These cell-based assays will be used to optimize antibody dependent immune responses directed against target cells. In the future the knowledge gained by these experiments and the techniques developed will be valuable in developing antibody-based protein therapeutics directed against cancer and other diseases.

Public Health Relevance

Antibodies are important therapeutic proteins that are used to treat cancers, arthritis, and many other diseases. The goal of this project is to understand how modification of human antibodies with sugars regulates antibody dependent immune responses, and then use the knowledge gained from these studies to develop better antibody-based treatments for diseases.

National Institute of Health (NIH)
National Institute of General Medical Sciences (NIGMS)
Research Project (R01)
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Macromolecular Structure and Function B Study Section (MSFB)
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Marino, Pamela
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Indiana University Bloomington
Schools of Arts and Sciences
United States
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More, Apurva S; Toth 4th, Ronald T; Okbazghi, Solomon Z et al. (2018) Impact of Glycosylation on the Local Backbone Flexibility of Well-Defined IgG1-Fc Glycoforms Using Hydrogen Exchange-Mass Spectrometry. J Pharm Sci 107:2315-2324
White, Derek R; Khedri, Zahra; Kiptoo, Paul et al. (2017) Synthesis of a Bifunctional Peptide Inhibitor-IgG1 Fc Fusion That Suppresses Experimental Autoimmune Encephalomyelitis. Bioconjug Chem 28:1867-1877
Pisupati, Karthik; Tian, Yuwei; Okbazghi, Solomon et al. (2017) A Multidimensional Analytical Comparison of Remicade and the Biosimilar Remsima. Anal Chem 89:4838-4846
Pisupati, Karthik; Benet, Alexander; Tian, Yuwei et al. (2017) Biosimilarity under stress: A forced degradation study of Remicade® and Remsima™. MAbs 9:1197-1209
Shah, Ishan S; Lovell, Scott; Mehzabeen, Nurjahan et al. (2017) Structural characterization of the Man5 glycoform of human IgG3 Fc. Mol Immunol 92:28-37
Lakbub, Jude C; Clark, Daniel F; Shah, Ishan S et al. (2016) Disulfide Bond Characterization of Endogenous IgG3 Monoclonal Antibodies Using LC-MS: An Investigation of IgG3 Disulfide-mediated Isoforms. Anal Methods 8:6046-6055
Okbazghi, Solomon Z; More, Apurva S; White, Derek R et al. (2016) Production, Characterization, and Biological Evaluation of Well-Defined IgG1 Fc Glycoforms as a Model System for Biosimilarity Analysis. J Pharm Sci 105:559-574
Mozziconacci, Olivier; Okbazghi, Solomon; More, Apurva S et al. (2016) Comparative Evaluation of the Chemical Stability of 4 Well-Defined Immunoglobulin G1-Fc Glycoforms. J Pharm Sci 105:575-587
Tauzin, Alexandra S; Laville, Elisabeth; Xiao, Yao et al. (2016) Functional characterization of a gene locus from an uncultured gut Bacteroides conferring xylo-oligosaccharides utilization to Escherichia coli. Mol Microbiol 102:579-592
Cuskin, Fiona; Lowe, Elisabeth C; Temple, Max J et al. (2015) Human gut Bacteroidetes can utilize yeast mannan through a selfish mechanism. Nature 517:165-169

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