# Method for the validation by Western analysis of affinity reagents against post-translationally modified proteins.

> **NIH NIH R44** · ABBRATECH, INC. · 2024 · $1,121,971

## Abstract

ABSTRACT
Antibodies (Abs) are commonly employed in biomedical research to identify and quantify target
proteins. However, the specificity, sensitivity and reliability of these Abs can be variable, which is a
significant concern in the scientific community. The use of an incorrect Ab can result in misleading
or inaccurate results, impeding progress in various areas of biomedical research. In Phase I of this
project, we developed tools to validate the specificity of commercial Abs used to detect
phosphorylated sites in proteins in a method we developed and published that we named
MILKSHAKE. We will present some of these results from this Phase I effort, which demonstrates the
critical need to address this concern. This proposed SBIR Phase II project aims to both extend and
commercialize the Phase I effort. We will continue the Ab validation work initiated in Phase I by
extending the analysis to other types of post-translational modifications (PTMs) of proteins in
Western analysis and flow cytometry. This Phase II project also seeks to improve and scale this
pipeline further, and to develop new methods for validating Abs against glycosylated proteins, which
are often dynamic and challenging to detect using traditional techniques. We will also employ an
innovative tool for measuring Ab promiscuity using a technology we term ‘Sundae’ to better
investigate the specificity of the Ab paratope. Sundae will be demonstrated for two therapeutic Abs.
The Phase II project has several key objectives, including the development of new Ab-specific
assays and the establishment of a centralized database for sharing validated Ab data. The project
team comprises experts in Ab validation, proteomics, and molecular biology, and will collaborate
closely with our current and future industry partners and academic researchers to ensure our
success.

## Key facts

- **NIH application ID:** 10931606
- **Project number:** 5R44GM146473-03
- **Recipient organization:** ABBRATECH, INC.
- **Principal Investigator:** Michael P Weiner
- **Activity code:** R44 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $1,121,971
- **Award type:** 5
- **Project period:** 2022-04-01 → 2026-07-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10931606

## Citation

> US National Institutes of Health, RePORTER application 10931606, Method for the validation by Western analysis of affinity reagents against post-translationally modified proteins. (5R44GM146473-03). Retrieved via AI Analytics 2026-05-26 from https://api.ai-analytics.org/grant/nih/10931606. Licensed CC0.

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