Protein profiling, quantitative

Von Haller, P.D., Yi, E., Donohoe, S., Vaughn, K., Keller, A., Nesvizhskii, A.I., Eng, J., Li, X.J., Goodlett, D.R., Aebersold, R., Watts, J.D. (2003). The Application of New Software Tools to Quantitative Protein Profiling Via Isotope-coded Affinity Tag (ICAT) and Tandem Mass Spectrometry II. Evaluation of Tandem Mass Spectrometry Methodologies for Large-Scale Protein Analysis, and the Application of Statistical Tools for Data Analysis and Interpretation. Mol. Cell. Proteomics 2, 428 -42. 

Smolka M, Zhou H, Aebersold R. Quantitative protein profiling using two-dimensional gel electrophoresis, isotope-coded affinity tag labeling, and mass spectrometry. Mol Cell Proteomics 2002 1 19-29. 

Figure 4-22. Densitometric analysis of serum separation. When electrophoresis is complete, the cellulose acetate film is removed and stained. Excess dye, not bound to protein, is removed by destaining and the film is transferred to a bath in which the cellulose acetate becomes transparent. The film is placed in a scanner in which it is moved across the light path of a spectrophotometer recording the absorbance of the blue stained protein peaks. The result is a densitometric scan whose peaks can be integrated to quantitate the protein profile of the serum.

Prokai, L., Zharikova, A.D. and Stevens, S.M. Jr. (2005) Effect of chronic morphine exposure on the synaptic plasma-membrane subproteomeofrats a quantitative protein profiling study based on isotope-coded affinity tags and liquid chromatography/mass spectrometry. J. Mass Spectrom. 40, 169-175. 

For nontargeted quantitative proteomics, in which the protein profiling and relative quantitation are performed simultaneously, mass spectrometers with high accuracy and resolution such as Q-TOF, TOF-TOF, or Orbitrap are... 

The most mature approach for quantitative protein profiling is two-dimensional gel electrophoresis (2DE). In 2DE, the sample under investigation is fractioned and... 

The other approach to quantitative protein profiling is based on stable isotope labelling of proteins and peptides and automated tandem mass spectrometiy (MS/MS) (Gygi et al., 1999b). This method, termed isotope-coded affinity tag labelling or ICAT, has been shown to be robust, reproducible and amenable to high throughput automation (Ideker et al.,... 

Approaches aimed at achieving quantitative protein profiling using antibody arrays in a multiplex format include signal amplification, multicolor detection and competitive displacement assays (Barry and Soloviev, 2004). If the use of antibodies for protein microarrays is to be materialized, several important criteria have to be fulfilled ... 

The separation of dairy proteins by CE has been generally carried out by CZE and has been exhaustively covered in several review papers, - - thus Table 30.8 only presents the key methodologies that offer the reader an overview of their most distinctive features. Basically, dairy protein analysis has been performed in whole milk for the simultaneous determination of caseins and whey proteins, or in fractions isolated from milk after casein precipitation. The first approach being used when the quantitative determination of the major proteins is required for the calculation of casein/whey protein ratios or for authentication purposes where an analysis of the whole protein profile is required. In both cases, accurate quantitative data must be derived. However, few studies have addressed the analysis of both groups of proteins in a single run by presenting quantitative data based on calibration curves constructed with analytical standards and good recovery of all proteins from milk samples. 

To optimize milk protein production, requirements must be accurately determined and matched with dietary supply. Through microbial digestion in the reticulo-rumen, ruminants have a capacity to utilize forages that are indigestible in non-ruminants. Dietary proteins are degraded in the rumen and used in microbial protein synthesis, which modifies dietary supply of protein both quantitatively and qualitatively making predictions of the amount and profile of amino acids (AA) absorbed from the small intestine difficult. 

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