Key Points
- A new NAD+ reading technique called bioluminescence resonance energy transfer (BRET) only requires 5 µL of blood and takes readings with accuracy comparable to less efficient and more costly conventional methods.
- The new finger prick method verifies that NAD+ levels significantly fall for aged individuals between 50 and 80 years old.
- The new NAD+ measuring technique shows that although 500 mg doses of oral NMN for 30 days almost doubles blood NAD+, 1,000 mg doses of NMN increase blood NAD+ a further 30%.
Declining NAD+ levels with age in human blood, muscle, and saliva has been linked to aging and age-related conditions like frailty, arthritis, and heart failure. To avoid ailments associated with falling NAD+ levels during aging, researchers have proposed replenishing NAD+ with precursors like NMN. Conventional techniques that measure how NMN affects NAD+ levels like liquid chromatography-mass spectrometry (LC-MS) require a laboratory and multi-step sample preparations for analyses and so aren’t efficient or cost-effective. Coming up with more affordable and easy-to-apply NAD+ measurement methods will make large scale, population-based studies of NAD+ levels and replenishment techniques easier.
Published in Aging Cell, Yu and colleagues from the Shenzhen Institute of Advanced Technology Chinese Academy of Sciences reveal a new, more cost-effective NAD+ measurement technique that requires only 5 µL of blood. With this new technique, the researchers confirm that, on average, human blood NAD+ levels decline with age. Making use of their new technique, the researchers also show that 1,000 mg of NMN increases blood NAD+ significantly more than 500 mg. The study highlights a new, efficient and cost-effective way to measure human NAD+ and demonstrates its capabilities with experiments confirming NAD+ declines with age along with NMN’s effects on blood NAD+.
The Finger Prick Method to Measure NAD+ Confirms Declining NAD+ Levels and Optimal NMN Doses
Utilizing a new technique called BRET, based on NAD+ sensing proteins from the bacteria Escherichia coli that change conformation when bound to NAD+, Yu and colleagues collected 5 µL blood samples from 75 female and 67 male study participants. The researchers compared blood NAD+ readings between the BRET method and one of the conventional NAD+ reading techniques, LC-MS. Intriguingly, the NAD+ readings almost perfectly aligned between the two techniques. These results suggest that the new BRET technique can reliably read blood NAD+ levels, much like LC-MS.
To apply the accurate NAD+ reading BRET technique, Yu and colleagues sought to find whether it confirms that human blood NAD+ levels fall with age. The researchers split the female and male participants up by age and found that for both sexes, average NAD+ levels significantly declined in aged individuals compared to younger ones. These results confirmed that NAD+ levels decline with age in both sexes using the BRET technique.
With the utility of the BRET technique in reading blood NAD+ levels established, Yu and colleagues sought to apply the method to find whether 1,000 mg doses of NMN exceeded 500 mg doses in increasing blood NAD+. The researchers provided study participants with either dosage orally for 30 days. Interestingly, even though 500 mg doses of NMN almost doubled blood NAD+, 1,000 mg doses further elevated blood NAD+ by about 30%. These data suggest that the optimal NMN dose for increasing blood NAD+ is higher than 500 mg, possibly being closer to 1,000 mg.
“We demonstrated a low-cost and easy-to-use assay using recombinant sensor protein and an automated optical reader for measuring fingertip blood NAD+,” said Yu and colleagues.
Using the NAD+ Measurement Technique to Research Ways for Individually Tailored Anti-Aging Regimens
The study sheds light on BRET, a new, cost-effective way to measure human NAD+ levels. The method measures blood NAD+ with the accuracy of more expensive and less efficient techniques like LC-MS. It also confirmed that blood NAD+ levels decline as people grow older and that 1,000 mg doses of NMN surpass 500 mg doses for replenishing blood NAD+. Interestingly, the prominent Harvard Professor and NMN researcher David Sinclair takes 1,000 mg of NMN per day, which aligns with the study’s findings that 1,000 mg dosages surpass 500 mg for boosting NAD+.
Due to its low cost and ease of application, researchers can apply the BRET technique to more individuals to find if some people produce more or less NAD+ than average. Along those lines, identifying these subgroups with NAD+ measurements like BRET in tandem with future genetic analyses could lead to individually tailored anti-aging regimens that include supplementation with molecules like NMN. With the ease of BRET’s applicability, more discoveries related to NAD+ biology along with individualized age intervention treatment options related to increasing NAD+ could await in the next two to three years.
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