RAS-Fingerprint™: Angiotensin Quantification

Analysis

Angiotensins

Steroids

Enzyme Activity

Renin

Angiotensinogen

More 

Species

Human

Mouse

Rat

Dog

Pig

Sheep

Horse

Cat

Monkey

Sample Type

Blood

Serum

Plasma

Urine

CSF

Cell Culture

Kidney

Heart

Brain

Lung

Liver

Mass Spectrometry

Attoquant Diagnostics is the unique provider of mass spectrometry based angiotensin quantification.

We use mass spectrometry for all our assays due to its unchallenged accuracy to discriminate multiple peptides and its high sensitivity to detect hormones in the low picomolar molecular range of biological samples.
The combination of ultra-pressure-liquid chromatography-tandem mass spectrometry (LC-MS/MS) and stable-isotope-labeled internal standards provides unchallenged specificity and sensitivity in all our assays and is the backbone of our service.

Enzyme Assays

In order to assay Neprilysin, for instance, we determine the turnover of Ang I to Ang 1-7 in presence and absence of a specific Neprilysin inhibitor.

The measurement of protease activities in tissue and plasma samples is a challenging task as the importance of assuring signal specificity is widely underestimated. Common strategies aiming at determining the activity of proteolytic enzymes in complex samples employ the cleavage of artificial substrates.  Colored or fluorescent products that are released from a quenched artificial substrate upon protease cleavage are measured and enzyme activities are quantified using a standard curve of recombinant enzyme that is usually prepared in matrix free buffers. Importantly, biological samples contain dozens of proteases that can cleave such artificial substrates, which usually contain very few amino acids that aim at mediating the selectivity for a certain enzyme. As a consequence, especially low abundance enzymes cannot be accurately quantified in complex samples using commercially available colorimetric or fluorimetric kits.

 

Our LC-MS/MS based approaches combine the conversion of natural substrates into the natural product of a certain enzymatic reaction with the highly sensitive and direct quantification of the enzyme specific product. The additionally employed specific inhibitors guarantee the selectivity of the assay. Standard curves of recombinant enzymes are routinely prepared in the original sample matrix.

Assay Development

Based on our proprietary technology for mass spectrometry sample preparation we are able to offer a customized assay development program for low abundance peptides in plasma and tissues. We have broad experience in analyte enrichment from various biological samples and offer low budget feasibility studies for specialized analytic questions.