# Development of a biparatopic-domain antibody screening system

> **NIH NIH R21** · UNIVERSITY OF PITTSBURGH AT PITTSBURGH · 2024 · $233,898

## Abstract

The mature form of human mesothelin (MSLN) is a cell-surface protein that is normally expressed at a low
level on the mesothelial cells but is overexpressed in many cancers, including lung adenocarcinoma and
mesothelioma, making it a practical target for therapeutic antibody applications. Our long-term goal, to
develop MSLN-targeting domain antibodies that will be utilized to design therapeutic reagents. MSLN is
highly relevant to environmentally caused disease and human health. In this research, we propose to
develop a screening system for biparatopic human heavy chain (VH) domains in the tandem VH1-VH1- form,
to find a better MSLN binder and to test whether biparatopic screening will identify novel binders compared
to tethered VH domains generated using individually screened VHs. In Aim 1, we will identify binding sites of
the tethered VH1-VH2- that were generated using the individually screened VHs. In Aim 2, we will develop a
direct screening system of the biparatopic antibody domain in the VH1-VH2- form. Results obtained in each
Aim will be independently fruitful to develop MSLN antibodies. We will also compare VH1-VH2 sequences
and MSLN-binding sites obtained in Aim 2 with those in Aim 1. By identifying how the sequences and
binding site are changed by the screening in Aim 2, we will scale up anti-MSLN VH1-VH2 screening and their
characterization, including optimization of biparatopic CAR-T cell design, immunoncology and animal
studies, and [in the future] plan to generalize the biparatopic domain antibody screening approach to other
important tumor-associated antigens.

## Key facts

- **NIH application ID:** 10789443
- **Project number:** 1R21ES035885-01
- **Recipient organization:** UNIVERSITY OF PITTSBURGH AT PITTSBURGH
- **Principal Investigator:** RIEKO ISHIMA
- **Activity code:** R21 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $233,898
- **Award type:** 1
- **Project period:** 2024-01-18 → 2025-12-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10789443

## Citation

> US National Institutes of Health, RePORTER application 10789443, Development of a biparatopic-domain antibody screening system (1R21ES035885-01). Retrieved via AI Analytics 2026-05-25 from https://api.ai-analytics.org/grant/nih/10789443. Licensed CC0.

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