Kuwait University Recombinant Therapeutic Proteins II PDF

1120-111 Biochemistry and Biotechnology Fundamentals Recombinant Therapeutic Proteins II 1120-111 Biochemistry and...

1120-111 Biochemistry and Biotechnology Fundamentals Recombinant Therapeutic Proteins II 1120-111 Biochemistry and Biotechnology Fundamentals Recombinant Therapeutic Proteins II  1120-111 Biochemistry and Biotechnology Fundamentals Recombinant Contents Therapeutic Proteins II Introduction to Anemia and Erythropoiesis Erythropoiesis-Stimulating Agents (ESAs) Overview of Interferons Recombinant Production Techniques Summary and Clinical Applications Recombinant Protein Instability  1120-111 Biochemistry and Learning Objectives Biotechnology Fundamentals Recombinant By the end of this lecture, you should be able to: Therapeutic Proteins II Explain the concept of anemia and the role of erythropoietin (EPO) and erythropoiesis-stimulating agents (ESAs) in managing anemia-related conditions. Describe the functions and types of interferons, including their natural roles in the immune system and FDA-approved therapeutic applications for various diseases. Discuss the recombinant production methods for proteins such as interferons and erythropoiesis- stimulating agents, including the use of genetically engineered cells like Escherichia coli and CHO cells. Recognize the clinical applications and FDA-approved uses of recombinant proteins, including ESAs for anemia treatment and interferons for viral infections, multiple sclerosis, and genetic immune disorders. Identify the main chemical and physical factors that contribute to the instability of recombinant proteins, including specific mechanisms like hydrolysis, oxidation, and denaturation.  1120-111 Biochemistry and Biotechnology Fundamentals Anemia Recombinant Therapeutic Proteins II Anemia is a problem of not having enough healthy red blood cells or hemoglobin to carry oxygen to the body's tissues. Having anemia can cause tiredness, weakness and shortness of breath. Anemia may be a sign of a more serious condition, such as bleeding in your stomach, inflammation from an infection, kidney disease, cancer, or autoimmune diseases.  1120-111 Biochemistry and Biotechnology Erythropoietin (EPO) Fundamentals Recombinant Therapeutic Proteins II Erythropoietin is a hormone (glycoprotein) that your kidneys naturally make to stimulate the production of red blood cells. High or low levels of erythropoietin can cause health problems. A healthcare provider can measure your erythropoietin levels with a blood test.  1120-111 Biochemistry and Biotechnology Fundamentals Recombinant Therapeutic Proteins II  1120-111 Biochemistry and Erythropoiesis stimulating agents Biotechnology Fundamentals Recombinant Recombinant Erythropoietin (EPO) Therapeutic Proteins II Erythropoiesis stimulating agents (ESAs) are recombinant forms of EPO produced synthetically via recombinant DNA technology in cell cultures. Examples of erythropoiesis-stimulating agents include epoetin alfa, darbepoetin. ESAs are generally indicated for patients with conditions associated with impaired red blood cell production. The 2 primary FDA-approved indications for ESA administration are: 1) anemia secondary to chronic kidney disease and 2) chemotherapy-induced anemia in patients with cancer. The FDA approved the use of epoetin (1993) and darbepoetin (2002) for patients with chemotherapy-induced anemia. The FDA has also approved ESAs for the treatment of anemia secondary to zidovudine therapy for HIV infection.  1120-111 Biochemistry and Erythropoiesis stimulating agents Biotechnology Fundamentals Recombinant Recombinant Erythropoietin (EPO) Therapeutic Proteins II During large-scale manufacturing protocols, erythropoiesis-stimulating agents are typically sourced from transfected Chinese hamster ovary cells (CHOs). An isotonic solution buffers the ESA powder, which the provider can administer intravenously or subcutaneously.  1120-111 Biochemistry and Biotechnology Fundamentals Interferon Recombinant Therapeutic Proteins II A natural substance that helps the body's immune system fight infection and other diseases, such as cancer. Interferons are made in the body by white blood cells and other cells, but they can also be made in the laboratory to use as treatments for different diseases. They are proteins made and released by host cells in response to the presence of pathogens such as viruses, bacteria, parasites or tumor cells. They allow for communication between cells to trigger the protective defenses of the immune system that eradicate pathogens or tumors. There are three types of interferons, namely interferon alpha, beta, and gamma Recombinant interferon: A process to isolate genetic material (DNA) containing the nucleotide sequence coding for interferon. Interferon alfa-2a is a recombinant DNA-derived protein prepared from genetically engineered Escherichia coli containing the human interferon α-2a gene. This protein is highly purified and has an antiviral activity.  1120-111 Biochemistry and Biotechnology Fundamentals Types Recombinant Therapeutic Proteins II Interferon-alpha is FDA-approved to treat chronic hepatitis C and B, and cell leukemia in adults. Interferon-beta is FDA approved for the treatment of multiple sclerosis (is a chronic disease of the central nervous system. MS is unpredictable. Some people may be only mildly affected. Others may lose the ability to see clearly, write, speak, or walk. Early symptoms can include vision problems, trouble walking, and tingling feelings.) IFN-γ is FDA approved to treat chronic granulomatous disease (a genetic disorder in which white blood cells called phagocytes are unable to kill certain types of bacteria and fungi) and osteoporosis.  1120-111 Biochemistry and Biotechnology Fundamentals Instability of recombinant Recombinant Therapeutic Proteins II proteins Loss of biologic activity may occur as a result of: Chemical Instability: results in bond formation or cleavage yielding a modified protein and a new chemical entity. Physical instability: involves a change in the secondary or higher-order structure of the protein.  1120-111 Biochemistry and Biotechnology Fundamentals Chemical Instability Recombinant Therapeutic Proteins II a) Hydrolysis: Peptide linkage between Asp-Pro is the least stable.  1120-111 Biochemistry and Biotechnology Fundamentals Recombinant Therapeutic Proteins II b) Hydrolytic deamidation: Asparagine and Glutamine are susceptible to deamidation to aspartate and glutamate.  1120-111 Biochemistry and Biotechnology Fundamentals Recombinant Therapeutic Proteins II c) Oxidations: Can occur at the side chains of sulfur-containing Methionine (Met) and cysteine (Cys).  1120-111 Biochemistry and Biotechnology Fundamentals Recombinant Therapeutic Proteins II d) Racemization: Inversion of configuration at asymmetric center, to yield residues in protein with mixtures of L and D configurations. This alters the protein physicochemical properties and biological activity.  1120-111 Biochemistry and Biotechnology Fundamentals Physical Instability Recombinant Therapeutic Proteins II May result from: - Denaturation - Non-covalent self aggregation (precipitation) Denaturation can be induced by: - Elevated temperatures - Drastic changes in pH - Addition of organic solvents  1120-111 Biochemistry and Biotechnology Fundamentals Recombinant Therapeutic Proteins II