# Proteome Scale Multiplexed Generation of Recombinant Antibodies

> **NIH NIH DP2** · UNIVERSITY OF OREGON · 2021 · $1,295,675

## Abstract

Project Summary/Abstract.
Antibodies are an invaluable tool in the biological sciences and are increasingly used as therapeutics. Over the
past few years a large number of proteomic techniques and large scale studies have been developed to probe
biological and biochemical processes at ever increasing scales. Many of these methods have been limited by
the availability of quality antibody reagents which are still generated one-by-one for each antigen target and the
lack of antibody sequences available. Additionally, large concerns have been raised regarding the quality and
reproducibility of current commercial antibodies. While high-throughput methods have been described capable
of generating antibodies against hundreds of antigens, they are slow, laborious, usually require significant
automation, and are dependent on antigen availability. These issues, combined with the growing need for
many quality antibodies, highlight the urgent need to develop methods capable of proteome-scale antibody
generation.
Here I introduce a radical proposal to create a platform which can generate antibodies against all potential
antigen targets in a target proteome and subsequently carry out proteome-scale tests of cross-reactivity.
This approach will be demonstrated on the human and zebrafish proteomes and consists of three parts.
First, I will develop gene synthesis technologies to create proteome-scale gene libraries consisting of tens of
thousands of antigens. Second, I will generate high diversity libraries of antibodies by repurposing diversity
generating retroelements to carry out targeted mutagenesis in-vivo. This will allow us to bypass the diversity
bottlenecks normally imposed by transformation. Third, I will carry out a large scale library-on-library selection
for antibody-antigen interactions in-vivo using a multiplexed protein fragment complementation assay. The
resulting antibody hits will be rescreened against all antigens to measure their cross-reactivity. This study will
reduce the cost of antibody generation by at least two-orders of magnitude and lead to the rapid creation of
large libraries of high-specificity, sequence-verified antibodies which, given the ubiquitous nature of these
reagents, will have a significant impact on all areas of basic and clinical research. Furthermore, the
technological developments in gene-synthesis and targeted in-vivo mutagenesis outlined will here have broad
applicability in many areas of life sciences research.

## Key facts

- **NIH application ID:** 10244823
- **Project number:** 1DP2TR004215-01
- **Recipient organization:** UNIVERSITY OF OREGON
- **Principal Investigator:** Calin Plesa
- **Activity code:** DP2 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $1,295,675
- **Award type:** 1
- **Project period:** 2021-09-10 → 2024-05-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10244823

## Citation

> US National Institutes of Health, RePORTER application 10244823, Proteome Scale Multiplexed Generation of Recombinant Antibodies (1DP2TR004215-01). Retrieved via AI Analytics 2026-05-22 from https://api.ai-analytics.org/grant/nih/10244823. Licensed CC0.

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