Neuropeptide reporter assay for serum, capillary blood and blood cards

U Schreiber,C Engl,M Bayer,S König

doi:10.1016/j.mex.2020.100985

Abstract

In the associated main paper (JPBA 2019), substance P was shown to be a valuable neuropeptide reporter substance to monitor the protease activity of serum. The assay was developed based on the predecessor assay using bradykinin (JPBA 2017). Both neuropeptides are of interest in inflammation and pain research and were thus explored for use with capillary blood and blood cards. Here, we present the protocols and set them in perspective to above neuropeptide assays for serum.•Neuropeptide reporter substance protease activity assay for use with fresh and dried blood.•Dabsylated Substance P and bradykinin are substrates of angiotensin-converting enzyme and other proteases.•Neuropeptides of interest in inflammation and pain.

Highlights

This MethodX article supplements work were we show the use of dabsylated substance P (DSP) as a neuropeptide reporter substance (NRS) for the monitoring of protease activity (PA) in serum [1]
This article is based on the assay developed for the use of dabsylated bradykinin (DBK) for the same purpose [2], which was already employed in a clinical study [3]
We have evaluated the application of the assay with respect to other types of sampling such as capillary blood and blood cards in order to extend the application range of the assay, which was only used with serum so far [1, 2]

Summary

Introduction

Biochemistry, Genetics and Molecular Biology Enzyme activity Neuropeptide reporter substance protease activity assay (NRS-PAA) Dabsyl-bradykinin protease activity assay (DBK-PAA) M. Dabsylated peptides were ordered from Peptide Specialty Laboratories (Heidelberg, Germany) Microcapillaries (Drummond Microcaps) allowed replicate collection with a defined volume (3 or 5 μl) Blood cards (Whatman 903 Protein Saver Card) This MethodX article supplements work were we show the use of dabsylated substance P (DSP) as a neuropeptide reporter substance (NRS) for the monitoring of protease activity (PA) in serum [1].

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Conclusion