Recombinant proteins engineered

Mattoussi H, Mauro JM, Goldman ER, Anderson CP, Sundar VC, Mikulec FV, Bawen-di MG (2000) Self-assembly of CdSe-ZnS quantum dot bioconjugates using an engineered recombinant protein. J Am Chem Soc 122 12142-12150... 

H. Mattoussi et ah, Self-Assembly of CdSe-ZnS Quantum Dot Bioconjugates Using an Engineered Recombinant Protein. J. Am. Chem. Soc., 122(49), 12142-12150 (2000). 

Stability and Purification of the Recombinant Protein. There are no hard and fast rules specifying, eg, whether a recombinant protein is available in a soluble state in the cell. In some cases, the expression system must be engineered by in vitro mutagenesis to optimize overall yield of the protein. 

The methods involved in the production of proteins in microbes are those of gene expression. Several plasmids for expression of proteins having affinity tails at the C- or N-terminus of the protein have been developed. These tails are usefiil in the isolation of recombinant proteins. Most of these vectors are commercially available along with the reagents that are necessary for protein purification. A majority of recombinant proteins that have been attempted have been produced in E. Coli (1). In most cases these recombinant proteins formed aggregates resulting in the formation of inclusion bodies. These inclusion bodies must be denatured and refolded to obtain active protein, and the affinity tails are usefiil in the purification of the protein. Some of the methods described herein involve identification of functional domains in proteins (see also Protein engineering). 

Ito, H., Steplewski, A., Alabyeva, T., and Fertala, A., Testing the utility of rationally engineered recombinant collagen-hke proteins for appheations in tissue engineering, J. Biomed. Mater. Res. A., 76 (3), 551-560, 2006. 

Smith, M. C., Engineering metal binding sites into recombinant proteins for facile purification, Ann. N. Y. Acad. Sci., 646, 315, 1991. 

A final problem for bioinformatics and bioanalytical scientists is the characterization of engineered microorganisms. Whole-cell analysis by mass spectrometry has been used to confirm the introduction of therapeutic genes into adenovirus vectors,100 to confirm the expression of recombinant proteins in bacteria,101,102 and also in vaccinology.103 In the broader case, identification of... 

Choi, D.H. and Keum, K.C. (2006) Production of recombinant proteins by high cell density culture of Pichia pastoris. Chemical Engineering Science, 61, 876-885. 

Several areas are receiving much of the research attention. Approaches that integrate product recovery with the fermentation in a three-phase fluidized bed bioreactor reflect general research trends in biochemical engineering (Yabannavar and Wang, 1991 Davison and Thompson, 1992). The successful use of three-phase biofluidization has also been demonstrated for recombinant protein systems, where it may have some benefit in improving plasmid stability (Shu and Yang, 1996). 

The Field Evaluation of Transgenic Crops Engineered to Produce Recombinant Proteins... 

Many of the initial biopharmaceuticals approved were simple replacement proteins (e.g. blood factors and human insulin). The ability to alter the amino acid sequence of a protein logically coupled to an increased understanding of the relationship between protein structure and function (Chapters 2 and 3) has facilitated the more recent introduction of several engineered therapeutic proteins (Table 1.3). Thus far, the vast majority of approved recombinant proteins have been produced in the bacterium E. coli, the yeast S. cerevisiae or in animal cell lines (most notably Chinese hamster ovary (CHO) cells or baby hamster kidney (BHK) cells. These production systems are discussed in Chapter 5. 

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