Temperature jump DNP

Dissolution DNP and temperature jump DNP are two related concepts that share the common principle of carrying out the DNP enhancement at low temperatores. 

To study rates of antibody-hapten reactions the principal methods employed have involved stopped flow or temperature jump techniques. The former was first used by Sturtevant et al. (64) and Day et al. (65). The temperature jump method has been employed by Froese, Sehon, and their collaborators (66-68) and more recently by Pecht et al. (69). Both methods are utilized in conjunction with very rapid optical measurements (in the millisecond range). For example, Sturtevant et al. took advantage of a spectral shift which occurs upon combination of anti-Dnp antibody with the dye, 2-(Dnp-azo)-I-naphthol-3,6-disulfonic acid (64). With the same hapten, and with e-Dnp-L-lysine and e-Dnp-6-aminocaproate. Day et al. (65) used the method of fluorescence quenching (Section VI,D) with a stopped flow apparatus. In the temperature jump technique the components are first equilibrated, a temperature increment is rapidly induced (up to 10°C in 0.1 isecond), and the rate of reequilibration at the new temperature is measured. Velocity constants can be estimated from the data the mathematical approaches required are described in the references cited. 

Of particular interest is the use of uv-visible spectrophotometry, since there is the added possibility of following the kinetics of simple hydrogen-bond formation by means of the temperature-jump relaxation technique with a conventional optical detection system. The method is suitable for acid or base species which contain a suitable chromophore, for example 2,4-dinitrophenol (2,4-DNP),... 

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