Protein and DNA Analysis

Protein digestion refers to the method in which the protein of interest is first digested by a proteolytic enzyme and then analyzed by mass spectrometry. This process affords a peptide map that is unique for each protein and allows identification 

Besides the benefits of scale reduction and trypsin immobilization, microsystem technology has other advantages to offer. For example, Ekstrom et al. [345] described a device that integrated an enzyme microreactor with a sample pretreatment robot and 

A heterogeneous assay was developed by Kartalov and Quake for DNA sequencing-by-synthesis to reliably sequence up to four consecutive base pairs [374], Briefly, the 

Digestion of proteins Trypsin Monolithic capillary column [348] 

High-speed online Trypsin, Porous silicon [353] 

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Soft ionization methods produce few fragments under relatively mild conditions. The ionization method that has received the most attention in terms of its applicability to protein and DNA analysis is the electrospray ionization (ES) technique. This is a soft method that is capable of generating molecular ions from biological macromolecules present in solution. Table 12.3 gives examples of the charge and m/z ranges that have been observed with some biopolymer species in electrospray ionization mass spectrometers. 

Rehhein H (1997). Fish as food fish species determination for caviar by protein and DNA analysis. Anim. Res. Dev., 46 39 16. 

Mass spectrometry is among the most powerful tools in protein and DNA analysis. It can determine molecular weights of biomolecules as large as 500,000 Da with high accuracy. Structural information like the amino acid sequence in a peptide or the sugar sequence in an oligosaccharide can be obtained. Some mass spectrometers can be coupled directly to a separation method such as LC or CE to combine the strengths of both techniques. 

Another direction in the development of electromigration techniques is the use of microchip technology. Microfabricated chips have advantages compared to conventional capillaries. They are disposable, rugged, and inexpensive. In addition, these systems can offer enhanced performance in terms of fast response and increased analysis speed. Through the use of chips with several separation microchannels, greater improvement in analysis speed is easily achieved. However, the applications of on-chip CE an on-chip CEC have centered on protein and DNA analysis-ti i i l... 

Matern D, Strauss AW, Hillman SL, Mayatepek E, Millington DS, Trefz FK (1999) Diagnosis of mitochondrial trifunctional protein deficiency in a blood spot from the newborn screening card by tandem mass spectrometry and DNA analysis. Pediatr Res 46 45-49... 

Karlin, S. and V. Brcndel "Chance and Sttnisiical Significance in Protein and DNA Sequence Analysis," Science. 39 (July 3. t992i,... 

Fig. 19.1. Tip-enhanced Raman (TERS) spectroscopic analysis of a single tobacco mosaic vims (TMV) particle (A) AFM image of the single virus and (B) TERS spectrum of the virus particle with an excitation wavelength of 568.2 nm showing contributions of protein and DNA...

Sulfur mustard (mustard gas) remains one of the CW agents of greatest concern because of its ease of production, favorable physicochemical properties, and potent vesicant action. It is a bifunctional alkylating agent, which reacts rapidly under physiological conditions with nucleophilic sites in proteins and DNA to form covalent adducts, via an intermediate episulfonium ion (see Figure 1). In the sections below, the various adducts (as unambiguously elucidated in recent years by mass spectrometry) are addressed, and methods for their analysis are discussed. 

Mass Spectrometric and Immunochemical Analysis of Covalent Adducts to Proteins and DNA. 433... 

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