# A Fluorescence-Based High-Throughput Platform for Glycotyping the Hematopoietic Cell Lineage

> **NIH NIH U01** · ALBERT EINSTEIN COLLEGE OF MEDICINE · 2021 · $427,520

## Abstract

PIs: Buelow – Steidl
 Project Summary
 The surface characteristics of cells in multicellular organisms are a direct reflection of their function.
Sensitive methods for detecting and defining novel surface characteristics of cells offer great opportunities for
gaining new mechanistic insights and developing new therapeutic strategies. Heparan sulfate (HS) glycans are
ubiquitous on the cell surface of metazoans and are expressed with extraordinary temporal and spatial
resolution. Since HS are amongst the most diverse molecules in nature, as a result of complex modification
patterns of the glycans, they could potentially provide the means to strongly discriminate between different cell
and tissue types. However, current analytical methods for HS glycan structure determination are expensive,
technically challenging and often low resolution, limiting their discriminatory power and widespread use. The
development of novel tools to define and exploit the structural characteristics of HS on the surface of cells
could open new avenues with important diagnostic and, possibly, therapeutic potential. We will develop a high-
throughput, multiplexed fluorescence-based platform to define the HS surface characteristics using a set of
HS-specific single chain variable fragment (scFvs) antibodies. We will first engineer the set of HS-specific
single-chain variable fragment antibodies with various immunotags as well as directly label them with
fluorescent tags using Sortase based approaches. Next, we will systematically determine the HS motifs that
are recognized by the scFv antibodies using a combination of microarrays with defined heparan sulfate
oligosaccharides and novel computational tools, which we will develop to define the HS motifs recognized by
these reagents. Finally, in proof of principle experiments, we will apply our tool box of immuno reagents against
defined HS motifs as an orthogonal system to the well-established cluster of differentiation (CD) system to
characterize and define hematopoietic cell lineages. We will leverage our laboratories' unique capabilities in
protein production and hematopoiesis research to generate, test, and disseminate the fluorescently labeled
scFvs reagents and protocols to the research community. We anticipate that our robust, efficient and
economical assays will lead to more widespread and standardized analyses of HS characteristics, which will
lay the foundation for the development of novel diagnostic and therapeutic approaches in the blood and other
systems in the future.

## Key facts

- **NIH application ID:** 10248374
- **Project number:** 5U01CA241981-03
- **Recipient organization:** ALBERT EINSTEIN COLLEGE OF MEDICINE
- **Principal Investigator:** Hannes Erich Buelow
- **Activity code:** U01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $427,520
- **Award type:** 5
- **Project period:** 2019-09-01 → 2023-08-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10248374

## Citation

> US National Institutes of Health, RePORTER application 10248374, A Fluorescence-Based High-Throughput Platform for Glycotyping the Hematopoietic Cell Lineage (5U01CA241981-03). Retrieved via AI Analytics 2026-05-24 from https://api.ai-analytics.org/grant/nih/10248374. Licensed CC0.

---

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