The analysis of mechanistic pathway markers and degradation products has led to the discovery of many scientific breakthroughs and novel therapeutic treatments.
For example, a robust analysis of peptides in the gastrointestinal (GI) tract has led scientists to relate these peptides to a range of other functional systems, including the cardiac, endocrine, and central nervous systems.
Linking these peptides is key to understanding how the body functions as an equilibrium. For example, the purification and identification of peptides in various systems is highly reliant on linking peptide relationships.
The G protein-coupled receptor (GPCR) has significantly developed new therapeutic treatments. While each of the analytes shown has different modes of action, each also interacts with the GI tract and binds to GPCRs.
Recent research has implicated the GI tract in the generation of diseases stemming from metabolic peptide dysfunction. By isolating these peptides, it may be possible to detect the metabolic changes occurring in cases where the cascade is not functioning correctly.
Pathway dysfunction facilitates a range of chronic diseases, including endocrine disorders such as diabetes or thyroid disease, kidney disease, and cell proliferation.
Hamilton Company’s portfolio of High-Performance Liquid Chromatography (HPLC) stationary phases is invaluable in isolating GPCR-binding peptides.
The example presented here analyzes seven peptides, highlighting the benefits of polymer-based reversed-phase resin PRP-3. Elevated flow rates significantly reduced analysis time, allowing for more rapid re-equilibration and sharper analyte peaks.
Covalent bonds present in PRP-3 also exhibit useful interactions between the peptides' biological π bonds and the resin’s available aromatic benzyl rings.
The resin is highly hydrophobic and comprises crosslinked divinylbenzene rings almost impervious to overtly alkaline or acidic mobile phases. This characteristic enabled excellent reproducibility between batches and multiple runs.
Source: Hamilton Company
| Column Information |
| Packing Material |
PRP-3, 7 µm |
| Dimensions |
150 x 4.6 mm |
| P/N |
79466 |
| Chromatographic Conditions |
| Gradient |
0.0–10.00 min. 15–30 % B |
| Temperature |
Ambient |
| Injection Volume |
5 µL |
| Detection |
UV at 215 nm |
| Eluent A |
H2O + 0.04 % HFBA |
| Eluent B |
CH3CN + 0.03 % HFBA |
| Flow Rate |
2.0 mL/min |
Compounds:
- Met-enkephalin
- Leu-enkephalin
- Neurotensin
- Bombesin
- Angiotensin II
- Somatostatin
- Insulin
Image Credit: Hamilton Company
Acknowledgments
Produced from materials originally authored by Adam L. Moore PhD. from Hamilton Company.
About Hamilton Company
Hamilton — The Measure of Excellence
Hamilton Company specializes in the development, manufacturing and customization of precision measurement devices, automated liquid handling workstations, and sample management systems. Hamilton's processes are optimized for quality and flexibility. Whether it's a custom needle with a quick delivery time frame, a special length pH sensor, or a comprehensive solution to fully automate your assay workflow, trust that Hamilton products will always meet your needs.
Hamilton Company has been a leading global manufacturer for more than 60 years, with headquarters in Reno, Nevada; Franklin, Massachusetts; Timișoara, Romania; Bonaduz, Switzerland; and subsidiary offices throughout the world.
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