Cell proteins analysis

Hoffman, K. (2000). Strategies for host cell protein analysis. Biopharm-Appl TBio 13(5) 38 15. 

Harwood MM, Christians ES, Fazal MA, Dovichi NJ. Single-cell protein analysis of a single mouse embryo by two-dimensional capillary electrophoresis. J Chromatogr A 2006 1130 190-4. 

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. 

Michels, D.A., Hu, S., Schoenherr, R.M., Eggertson, M.J., Dovichi, NJ. (2002). Fully automated two-dimensional capillary electrophoresis for high sensitivity protein analysis. Mol. Cell. Proteomics 1, 69-74. 

Zhang, Z., Krylov, S., Arriaga, E.A., Polakowski, R., Dovichi, N.J. (2000). One-dimensional protein analysis of an HT29 human colon adenocarcinoma cell. Anal. Chem. 72, 318-322. 

Joubert-Caron R et al. Protein analysis by mass spectrometry and sequence database searching a proteomic approach to identify human lymphoblastoid cell line proteins. Electrophoresis 2000 21 2566— 2575. 

A proteomic study of the HeLa cell proteins in raft fractions, identified by these criteria and estimated by quantitative mass spectrometry, has identified 241 authentic raft proteins [23]. This analysis found that the raft proteins ... 

Nunoi and co-workers (1988) fractionated neutrophil cytoplasm by Mono Q anion-exchange chromatography and obtained three fractions (NCF-1, -2 and -3) that were active in the assembly of the oxidase. Independently, Volpp and colleagues (Volpp, Nauseef Clark, 1988) prepared antiserum from cytosolic factors that eluted from a GTP-affinity column, and this antiserum (Bl) recognised cytoplasmic factors of relative molecular masses 47 kDa and 66 kDa. It was later shown by this group that these cytosolic factors translocated to the plasma membrane during activation. NCF-1 was shown to contain the 47-kDa protein and NCF-2 the 66-kDa protein. Analysis of the defect in the cytosol of autosomal recessive CGD patients revealed that most of these (88%) lacked the 47-kDa protein (p41 -phox), whereas the remainder lacked the 66-kDa protein (p66-phox). Both of these components have now been cloned and recombinant proteins expressed. Interestingly, in the cell-free system, recombinant p47-phox and p66-phox can restore oxidase activity of the cytosol from autosomal recessive CGD patients who lack these components. 

Los GV et al (2008) HatoTag a novel protein labeling technology for cell imaging and protein analysis. ACS Chem Biol 3 373-382... 

A study comprising five laboratories was carried out on the accuracy and precision of protein amino acid analysis. An important conclusion reached was that it is necessary to examine both accuracy and precision to achieve maximum improvement in either129. A commercially available single-cell protein, Pruteen, was proposed as reference material for the determination of amino acids and other substances in food. This recommendation was the result of a five-year-long study on the stability of this particular protein130. 

Our previous studies showed that CNTs exhibit marked cytotoxicity (Cui et al., 2005 Tian et al., 2006). For example, SWCNTs can inhibit HEK293 cell proliferation, decrease cell adhesive ability in a dose-and time-dependent manner as shown in Fig. 9.7. HEK293 cells also exhibit active responses to SWCNTs such as secretion of some 20-30 kd proteins to wrap SWCNTs, aggregation of cells attached by SWCNTs and formation of nodular structures as shown in Fig. 9.8. Cell cycle analysis showed that 25pg/ml SWCNTs in medium induced Gt arrest and cell apoptosis in HEK293 cells as shown in Fig. 9.9. Biochip analysis showed that SWCNTs can induce up-regulation expression of cell cycle-associated genes such... 

In this assay, rhuMAbs produced in transfected Chinese hamster ovary (CHO) cells were analyzed using a commercially available SDS-protein analysis... 

The Western blot method is often used in the analysis of host cell impurities. It can be used to identify a recurring impurity. O Keefe et al. used a Western blot to identify an E. coli protein impurity in the preparation of the recombinant fibroblast growth factor (aFGF).29 By using specific antisera to the E. coli host cell proteins, they were able to isolate the impurity and determine its N-terminus amino acid sequence to confirm its identity. Antibodies could be used to determine the concentration of this impurity in sample preparations. 

The compounds analyzed by GC/MS comprise e.g. amino acids [34, 39, 64 - 69], organic acids [33,63,65,66,69], sugars [39,70,71 ], lipids and fatty acids [72, 73]. Moreover, mass distributions of polymers and their building blocks, obtained via hydrolysis of the polymer, were assessed. Examples are glycogen [39, 70], cell protein [8,10,17], or DNA [74]. Most of the analytical methods have been developed for tissue samples. Since most of the compounds studied are polar or even charged molecules, derivatization is necessary in most of the cases of GC/MS analysis. The derivatization method of choice clearly depends... 

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