# Tailoring the Hydroxyl Radical Foot-Printing Approach to Provide a Solution for the Higher Order Structural Analysis Needs of the Biopharmaceutical Industry

> **NIH NIH R44** · GENNEXT TECHNOLOGIES, INC. · 2020 · $996,618

## Abstract

The GenNext Phase II SBIR submssion entitled “Tailoring the Hydroxy Radical
Foot-Printing Approach to Provide a Solution for the Higher Order Structural Analysis
Needs of the Biopharmaceutical Industry” is responsive to the ackowledged need for
new and improved tools for higher order structural analysis (HOS) of
biopharmaceuticals. Unlike conventional drugs, biopharmaceuticals are complex,
heterogeneous mixtures of 3-dimensional biomolecules, whose safety and efficacy is
reliant upon proper HOS. The presence of proteins with improper higher order structure
(HOS) has been linked to severe adverse drug reactions, alerting the biopharmaceutical
industry to the critical role of HOS, while establishing the need for new and improved
HOS analytics.
 An emerging HOS analysis technique is hydroxyl radical foot-printing (HRPF).
HRPF involves the irreversible labeling of a protein’s exterior by reaction with hydroxyl
radicals with subsequent MS analysis to identify the outer portions of the protein. The
most widely used method for generating OH radicals employs a quick burst of UV light,
and is appropriately called fast photochemical oxidation of proteins (FPOP). Typically, a
powerful and expensive UV laser is used. Academic laboratories have demonstrated the
utility of FPOP for HOS analysis; however adoption in pharma has been minuscule at
best. We have identified barriers that have limited the adoption of the HRPF approach in
the biopharmaceutical industry. These impediments include: 1) The use of expensive
lasers that demand substantial safety precautions; and 2) the irreproducibility of FPOP
caused by background scavenging of OH radicals that complicate and limit comparative
studies. As such, there are no commercial sample preparation devices for FPOP
analysis, despite the demonstrated need for the HOS analytical power.
 The GenNext proposal creates an improved means of performing HRPF analysis
by replacing expensive, hazardous lasers with a flash oxidation system. Moreover, a
custom internal standard (dosimetry) system will be included to facilitate ease of use and
improve reproducibility. These innovative advancements will decrease the barrier to
adoption of HRPF for HOS analysis and will result in accelerated adoption with
concomitant impact on biopharmaceutical research and development. In addition to
instrumentation sales, GenNext will offer a fee-for-service option for its clients who wish
to constrain upfront cost and risk, as they evaluate FOX HOS impact to their program.

## Key facts

- **NIH application ID:** 9902463
- **Project number:** 5R44GM125420-03
- **Recipient organization:** GENNEXT TECHNOLOGIES, INC.
- **Principal Investigator:** Scot Randy Weinberger
- **Activity code:** R44 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $996,618
- **Award type:** 5
- **Project period:** 2017-09-01 → 2021-12-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/9902463

## Citation

> US National Institutes of Health, RePORTER application 9902463, Tailoring the Hydroxyl Radical Foot-Printing Approach to Provide a Solution for the Higher Order Structural Analysis Needs of the Biopharmaceutical Industry (5R44GM125420-03). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/9902463. Licensed CC0.

---

*[NIH grants dataset](/datasets/nih-grants) · CC0 1.0*