# Activity-based discovery and optimization of agonist antibodies

> **NIH NIH F32** · UNIVERSITY OF MICHIGAN AT ANN ARBOR · 2021 · $69,704

## Abstract

PROJECT SUMMARY
Agonist antibodies, capable of initiating cellular signaling events through interaction at the cell surface, have
emerged as a new paradigm in disease treatment with several promising candidates in clinical trials including T
cell activating antibodies. However, the discovery of rare antibodies possessing specific biological functions
remains a major biotechnological bottleneck. To address this issue, I seek to employ direct function-based
antibody screening methods, an emergent biomedical technology using mammalian intracellular reporter
systems for which proof-of-concept has recently been shown for the discovery of agonist antibodies. I believe
that this innovative technology has the potential to be revolutionary if it can be generalized. Thus, the goal of
the proposed research is to establish new fundamental methods for broad use in function-based antibody
screening, which could ultimately lay the foundation for advancing disease treatment. I have recently
developed novel model systems that enable robust detection of intracellular signaling via the NF-κB pathway
initiated from extracellular activation of the T-cell receptors Ox40 and CD137. These receptors were chosen for
their availability of well-defined control agonist antibodies that are invaluable for system validation. I propose to
use these model systems to gain new understanding of the fundamental mechanisms and principles involved
in function-based antibody screening and employ this knowledge toward the development of new biomedical
technology and novel therapeutics via three distinct aims. First, I propose to critically evaluate the relationship
between biological activity, antibody display level, receptor display level, and antibody molecular format to
identify screening methods that ultimately improve the efficiency of agonist antibody discovery. Second, I
propose to develop new Ox40 and CD137 reporter cell systems for simplified and improved function-based
antibody discovery via genetic engineering of autocatalytic agonist antibody expression that depends on
receptor-specific intracellular signaling in order to improve agonist antibody discovery by both simplifying signal
detection and amplifying true positive signals. Thirdly, I seek to identify new Ox40 and CD137 agonist
antibodies with improved activity by direct function-based screening. Toward this goal, I will employ
computational approaches to inform library design in order to increase the likelihood that library variants
effectively engage T cell receptors near the active site. I will employ next-generation sequencing to identify
lead candidates, which will then be rigorously evaluated via classic T cell activation assays. Finally, I will
critically analyze lead sequences in relationship to agonist activity in order to uncover potential molecular
determinants of agonist activity and structure-function relationships. Overall, the proposed research has the
potential for broad therapeutic impact given that ins...

## Key facts

- **NIH application ID:** 10159090
- **Project number:** 5F32GM137513-02
- **Recipient organization:** UNIVERSITY OF MICHIGAN AT ANN ARBOR
- **Principal Investigator:** John Samuel Schardt
- **Activity code:** F32 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $69,704
- **Award type:** 5
- **Project period:** 2020-04-01 → 2022-04-15

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10159090

## Citation

> US National Institutes of Health, RePORTER application 10159090, Activity-based discovery and optimization of agonist antibodies (5F32GM137513-02). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10159090. Licensed CC0.

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