Affinity chromatography by Sameh Magdeldin

By Sameh Magdeldin

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Plasmids, nowadays mostly commercially prepared, are often used as suitable DNA vectors. Prepared expression plasmid is introduced into host cells. , 2001; Sørensen & Mortensen, 2005; Vodrážka, 1999). A scheme of the process of recombinant protein production is for a better comprehensibility shown in the figure 4. Another advantage offered by recombinant proteins production is the possibility of modification of the original nucleotide sequence encoding the protein of interest. For example suitable alterations in the original structure of the protein may result in desirable changes of its stability, activity or specificity.

Steps of a of reversed phase chromatography separation. 7. Funds This work was supported by JSPS (Japan Society for Promotion of Science) Grant-in-Aid for scientific research (B) to Dr. Sameh Magdeldin (23790933) from Ministry of Education, Culture, Sports, Science and Technology of Japan. The funders had no role in the decision to publish or in the preparation of this chapter. 8. References Allocati N, Federici L, Masulli M, Di Ilio C (2009) Glutathione transferases in bacteria. FEBS J 276: 58-75 Allocati N, Federici L, Masulli M, Di Ilio C (2011) Distribution of glutathione transferases in Gram-positive bacteria and Archaea.

G. immobilization of horseradish peroxidase via monoclonal enzyme specific antibody and anti-Fc antibodies) to preparation of fusion protein and its interaction with anti-tag antibody (used for alkaline phosphatase). e. Protein A from Staphylococcus aureus with the ability to specifically bind the Fc region of immunoglobulins and streptococcal Protein G with broader binding abilities. Both mentioned proteins enable oriented immobilization of antibodies on the surface, thus ensuring a correct orientation of antibody binding sites for further interaction.

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