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NeuCode – hero

NeuCode

NeuCode affords a different approach to protein quantification by exploiting the subtle mass differences of neutron binding in C, N, and H. At NCQBCS, we are developing the method to deliver up to 9-plex capacity. This technology is particularly exciting as it can offer multiplexed metabolic labeling, the gold standard method, for cell culture and in vivo labeling of mammals.

How NeuCode Works

How NeuCode Works

    

Consider a lysine molecule that has two additional neutrons – i.e., +2 Da. There are six ways one could build this molecule, e.g., 2 15N or 2 2H, and so on. Because of differences in neutron binding energy, each of these isotopologues has a distinct mass, with the two aforementioned examples differing by 18.5 mDa. Inclusion of four, eight, or 12 additional neutrons into the lysine molecules allows one to generate nearly a hundred unique isotopologues, all spaced about 1 mDa apart.


High resolution mass analysisorbitrap-web

High resolution mass analyzers, i.e., Fourier Transform instruments, especially the Orbitrap, have become widespread and offer the resolving power necessary to distinguish NeuCode lysine isotopologues embedded in identical peptides. Our initial description of NeuCode SILAC utilized two +8 Da heavy lysine amino acids, one with six 13C atoms and two 15N atoms and the other with eight 2H.[1] These two isotopologues differ in mass by 36 mDa and our experiments demonstrated that m/z peaks separated by this mass difference are easily distinguished at resolving powers above ~ 100,000

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When using NeuCode a low resolution MS1 scan is first acquired and used to select precursors for data-dependent MS/MS analysis. Next, a high resolution MS1 scan is acquired, in this case using an Orbitrap analyzer, permitting detection of the two NeuCode peaks and, thus, allowing quantification. While this scan is being acquired tandem mass spectra are taken using the ion trap analyzer on the Orbitrap ion trap hybrid system. Note that spectral complexity of the low resolution (R = 30,000) MS1 and the MS/MS scan are not impacted by the presence of the labels; in fact, you cannot even tell the labels exist unless they are revealed by high resolution analysis.

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Culturing Cells

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Culturing Cells

NeuCode is an ideal approach to perform multiplexed metabolic labeling for cultured cells. Currently up to seven plex labels are commercially available.  And, as instrument resolving powers continue to improve, NeuCode plexing capacity will likewise increase. Finally, the approach is ideal for primary cell lines where incomplete labeling is anticipated.  

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Multi-plexed in vivo Mouse Labeling

 

Multi-plexed in vivo
Mouse labeling

 NeuCode metabolic labeling can be extended to mammalian systems and offers a multiplexed method for adult mouse labeling and can be accomplished in as few as 10 days. This is the first method to permit such rapid in vivo labeling and eliminates the need to generate a labeled reference colony. 

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Technology – Training Link

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NeuCode Mouse

“This NeuCode chow is really tasty!”

              

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NeuCode Publication Library

Read more about other applications of NeuCode including its use for peptide and protein identification, top-down quantification, data-independent acquisition, and much more.

NeuCode Publication Library

Read more about other applications of NeuCode including its use for peptide and protein identification,
top-down quantification, data-independent acquisition, and much more.

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NeuCode Labeling for Top-Down Protein Quantification

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In Vivo Mouse Labeling in Just a Few Weeks Using NeuCode

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Multi-Plexed, Absolute Protein Quantification Using NeuCode

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NeuCode Offers a Route for Multi-Plexed Quantification with Data-Independent Acquisition 

 

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NeuCode Signatures Can be Used to Facilitate Peptide ID & De Noro Sequencing 

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Product Ion Annotation Using NeuCode

 

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A Simple Approach for NeuCode Chemical Labeling

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Neucode Labels for Metabolite Quantification

Industrial Partners

NeuCode is Available From Our Industrial Partners

Contact

For additional information about NCQBCS please send your contact details using the form below.
Note, if you are interested in training, please use the form on our training page.

Contact

For additional information about NCQBCS please send your contact details using the form below.
Note, if you are interested in training, please use the form on our training page.


Where we are

425 Henry Mall, Coon Lab 4462, Madison WI

777 Highland Avenue, Li Lab 5234, Madison, WI

330 N. Orchard Street, Pagliarini Lab 2268, Madison WI

 

How to contact us

Email address: laura.vantoll@wisc.edu