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A peptide immunoaffinity LC-MS/MS strategy for quantifying the GPCR protein, S1PR1 in human colon biopsies
Hongwei Zhang1, Eugene Ciccimaro1, Jacob Zalaznick1
1Research & Development, Bristol Myers Squibb, Princeton, NJ 08543, USA.
Abstract:
Background: S1PR1, a G protein-coupled receptor (GPCR) protein, is a therapeutic target for treatment of autoimmune diseases. As a potential biomarker for drug effect and patient stratification, it is of great significance to measure it in biological samples. However, due to the hydrophobic nature of S1PR1 and the difficulties in extraction and solubilization, as well as low expression levels, quantitative determination of S1PR1 remains challenging. Results: In this work, a peptide immunoaffinity LC-MS/MS method was developed to quantify S1PR1 in biopsy-sized colon samples with an LLOQ of 7.81 pM. Conclusion: Peptide immunoaffinity LC-MS/MS based strategy has achieved the desired sensitivity for low abundance S1PR1, and the same strategy could be applied to quantify S1PR1 in multiple species and other GPCR proteins.
Insights
Researchers developed a sensitive peptide immunoaffinity LC-MS/MS method to quantify S1PR1, a G protein-coupled receptor (GPCR), in colon samples. This breakthrough aids in measuring S1PR1 as a biomarker for autoimmune disease treatments.
Area of Science:
- Biochemistry
- Pharmacology
- Analytical Chemistry
Background:
- Sphingosine-1-phosphate receptor 1 (S1PR1) is a G protein-coupled receptor (GPCR) protein and a therapeutic target for autoimmune diseases.
- S1PR1 serves as a potential biomarker for assessing drug efficacy and stratifying patients in clinical settings.
- Quantifying S1PR1 is challenging due to its hydrophobic nature, low expression levels, and difficulties in extraction and solubilization.
Purpose of the Study:
- To develop a highly sensitive method for the quantitative determination of S1PR1 in biological samples.
- To overcome the challenges associated with measuring low-abundance, hydrophobic GPCRs.
Main Methods:
- Development of a peptide immunoaffinity liquid chromatography-tandem mass spectrometry (LC-MS/MS) assay.
- Application of the method to quantify S1PR1 in biopsy-sized colon samples.
Main Results:
- The developed LC-MS/MS method achieved a lower limit of quantification (LLOQ) of 7.81 pM for S1PR1.
- The peptide immunoaffinity strategy demonstrated high sensitivity for quantifying low-abundance S1PR1.
Conclusions:
- The peptide immunoaffinity LC-MS/MS strategy provides the necessary sensitivity for quantifying low-abundance S1PR1.
- This methodology is potentially applicable for S1PR1 quantification across different species and for other GPCR proteins.
