Biology subjects

Wiggins, C. D.

Publications and source records attributed to Wiggins, C. D..

3 recordsLinked to original sources

bioRxiv2026

An IgG-Optimized Enzyme-Linked Lectin Assay (ELLA) for Quantitative Analysis of Immunoglobulin Glycosylation

Antibody Fc glycosylation is modulated in a variety of disease and immune response contexts, altering downstream functional responses including antibody-dependent cellular cytotoxicity through modified immune cell Fc receptor binding. Accessible, high-throughput glycosylation assays such as enzyme-linked lectin assays (ELLAs) are essential to advance understanding of glycosylation regulation and function. However, current ELLA protocols lack standardization and optimization, and results are reported out in arbitrary absorbance units, limiting reproducibility and cross-study comparability. We developed an optimized multi-lectin parallel ELLA with three specific improvements: systematic optimization of incubation times and reagent concentrations; incorporation of Protein A for IgG specificity; and use of commercially available bovine fetuin B as a quantitative surrogate standard for cross-study reproducibility. Our panel of 8 lectins, SNA, RCA, LCA, PHA-E, PHA-L, MAL-I, WGA, and DSL, cover the major IgG glycoforms. We demonstrate that our ELLA panel can reveal biologically relevant cytokine-induced plasticity of IgG glycosylation profiles in immortalized B cells.

bioengineering↗

bioRxiv2025

Multivariate analysis of glycogenes reveals coordinated regulation of immunoglobulin glycosylation in an immortalized human B cell system

While neutralizing ability has traditionally been considered the most important antibody function, appreciation has grown for Fc-mediated extra-neutralizing functions, which are shaped by IgG glycosylation. However, there remain fundamental questions as to how B lymphocytes induce and regulate antibody glycosylation and thus functional capability. Understanding how transcriptional and cell state regulation shape glycosylation could reveal levers to tune protective humoral profiles in a disease- and antigen-specific manner. Prior studies have explored a limited panel of glycogenes and measured bulk glycosylation changes. Here, employing an in vitro antigen-specific B cell culture system, we systematically characterize transcriptional and humoral responses to cytokine perturbations. After exposure to a broad panel of cytokines (IL-4, IL-6, IL-10, IL-17, TNFa, IFNg, APRIL, and BAFF) across multiple concentrations and timepoints, transcriptomic profiling and lectin-based IgG glycome assays are employed to associate cytokine stimuli with both glycogene expression and IgG glycosylation. Supervised and unsupervised machine learning models identify cytokine-specific glycogene "signatures" as well as distinct immunoglobulin glycosylation profiles. We find that cytokines induce rapid transcriptional responses, with glycogene signatures outperforming single-gene changes in distinguishing stimulation conditions. We further demonstrate the ability to induce both pro- and anti-inflammatory IgG glycosylation profiles, particularly in terms of IgG galactosylation. This work demonstrates the utility of this system to parse the cytokine-driven regulation of B lymphocyte glycogenes, establishing a framework for dissecting how environmental cues shape antibody glycosylation, with relevance for autoimmune disease, infection, and vaccine responses.

systems biology↗

bioRxiv2023

Analysis of the Diverse Antigenic Landscape of the Malaria Invasion Protein RH5 Identifies a Potent Vaccine-Induced Human Public Antibody Clonotype

The highly conserved and essential Plasmodium falciparum reticulocyte-binding protein homolog 5 (PfRH5) has emerged as the leading target for vaccines that seek to protect against the disease-causing blood-stage of malaria. However, the features of the human vaccine-induced antibody response that confer highly potent inhibition of malaria parasite invasion into red blood cells are not well defined. Here we characterize over 200 human IgG monoclonal antibodies induced by the most advanced PfRH5 vaccine. We define the antigenic landscape of this molecule, and establish epitope specificity, antibody association rate and intra-PfRH5 antibody interactions are key determinants of functional anti-parasitic potency. In addition, we identify a germline gene combination that results in an exceptionally potent class of antibody and demonstrate its prophylactic potential to protect against P. falciparum parasite challenge in vivo. This comprehensive dataset provides a framework to guide rational design of next-generation vaccines and prophylactic antibodies to protect against blood-stage malaria.

immunology↗