**Author details**

348 Dehydrogenases

**Figure 19.** Synthesis of N6-carboxymethyl-NAD+

**O**

**P O P <sup>O</sup> OH**

**O OH**

**10. Conclusions** 

**N+**

**N N**

**2NH**

**NH2 O**

**N**

**N**

**R-ox**

**OH H H OH <sup>H</sup> <sup>H</sup> O**

The synthesized N6-carboxymethyl-NAD+ is an important intermediate for NAD immobilization at electrodes for continuous monitoring biosensors. An extension of this work could involve synthesis of various polymeric NADs with tailor-made chemical properties to meet biosensor requirements for continuous monitoring of different analytes.

**R-ox: R-red:**

**N**

**H H**

**NH2 O**

**OH H H OH <sup>H</sup> <sup>H</sup> O**

**OH <sup>H</sup> <sup>H</sup> OH <sup>H</sup> <sup>H</sup> <sup>O</sup> <sup>O</sup>**

**.**

**N N**

**NH**

**N**

**C OH O**

**N**

**R-ox**

**P O P <sup>O</sup> OH**

**O OH**

**O**

**N**

*1 23*

**2NH**

**C OH**

**O rearrangment**

**<sup>N</sup><sup>+</sup> <sup>N</sup>**

**N**

**R-red**

**enzyme reduction**

**OH <sup>H</sup> <sup>H</sup> OH <sup>H</sup> <sup>H</sup> <sup>O</sup> <sup>O</sup>**

**N**

**2NH**

**C OH**

**O iodoacetic acid**

**N+ <sup>N</sup>**

**.**

**N**

**R-ox**

In this article we have outlined various strategies for the electrochemical exploitation of dehydrogenase enzymes in sensor devices. The techniques used ultimately depend upon the class of dehydrogenase enzyme used. For enzymes that are NAD(P)+ linked it is essential to develop a base transducer (a modified electrode) that efficiently reoxidises the reduced coenzyme. Alternatively, sensing schemes can be designed which utilize Diaphorase thereby facilitating the biochemical oxidation of reduced coenzyme. With FAD and PQQ dependent enzymes the most successful strategy has been to utilize mediator molecules such as the ferricyanide anion to couple the enzymatic activity to the electrode. Although not yet exploited commercially, dehydrogenase enzymes could also have a role in continuous monitors. With the NAD+ dependent enzymes there is the additional complication of immobilizing the chemical components of the sensor to prevent drift in the device over time. This also includes the coenzyme molecule and we have illustrated how the synthesis of the N6-carboxymethyl derivative of NAD can be an important intermediate in achieving this. Attachment of suitable ligands at this position, with sufficient flexibility, should allow the development of stable reagents which will facilitate the development of continuous devices. The electrochemistry of NADH oxidation has been well researched over the last 30 years. Also, over 250 enzymes use this ubiquitous coenzyme so schemes which utilize NAD dependent enzymes should allow for the measurement of a range of analytes in blood. In spite of this, it is interesting to note that with the exception of Abbott, none of the other major biosensor manufacturers have embraced this technology. The reason for this could be due to the fact that glucose SMBG is still the largest biosensor market worldwide and it is adequately served by enzymes such as glucose oxidase, FAD-GDH and PQQ-GDH, none of Marco Cardosi and Zuifang Liu *LifeScan Scotland Limited, a Johnson & Johnson Company, UK* 
