Expression vectors therapeutic proteins

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... 

The desired gene/cDNA is normally amplified, sequenced and then introduced into an expression vector that facilitates its introduction and expression (transcription and translation) in an appropriate producer cell type. All recombinant therapeutic proteins approved to date are produced in E. coli, S. cerevisiae or in animal cell lines (mainly CHO or BHK cells). The general... 

Commercial Uses of Virus Expression Vectors 4.2.5.1 Vaccines and Therapeutic Proteins... 

Several plant viral vectors have been used to successfully produce vaccines and therapeutic proteins in plants. TMV-based expression vectors represent one of the more successful examples and have produced a wide array of therapeutic proteins such as a-trichosanthin, tumor-specific single-chain antibodies, and a number of vaccine antigens (Table 4.1) (Dalsgaard et al., 1997). More recent examples of plant viral expression vectors utilized for vaccine production are provided in detail in Koprowski and Yusibov (2001), Pogue et al. (2002), and Canizares et al. (2005). 

Selected Examples of Vaccines and Therapeutic Proteins Expressed in Plants Using Plant Viral Expression Vectors... 

The calli used to generate cell lines can be transgenic and express the vaccine or therapeutic protein of interest. Transfer of foreign genes into calli takes place by Agrobacterium-va.Q6M.td transformation, particle bombardment, electroporation of protoplasts, or by viral vectors (Fischer... 

Discusses the engineering of plant virus expression vectors for transient expression of vaccine proteins and other therapeutics in plant tissue... 

RNA to initiate cDNA synthesis. All cellular mRNA contains multiple repeats of adenine bases (poly-A tails). Therefore the complementary thymine bases (oligo-dT) can be used as a primer that binds to the mRNA template required for the reverse transcriptase to synthesize the cDNA. In the case of pancreatic mRNAs (Figure 4.2), the signihcantly higher mRNA for insulin compared with other proteins allowed success in isolating the insulin-specihc cDNA. Subsequent insertion of cDNA into a bacterial expression vector allowed the production of functional insulin that is now marketed as a successful therapeutic product (Figure 4.2). 

Early reports with rAd vectors may provide the proof of principle for transient expression of a number of potentially therapeutic proteins. For instance the Na+/K+-ATPase was found to promote reabsorption of alveolar fluid in an ARDS model (Factor et al., 1999 Sartori and Matthay, 2002). Anti-inflammatory cytokines and antioxidants have also been used with some success in a similar context in animal models of sepsis and ARDS (Van Laethem et al., 1998 Dumasius et al., 2002). 

This section focuses on the generation of engineered mammalian cell lines that stably produce therapeutic proteins. Hybridomas, transient expression systems, and insect and bacterial cell line development are outside the scope of this section. The commonly used cell lines, expression systems and vectors, as well as cell banking and stability are described. 

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