Recombinant Protein Production Flashcards
(29 cards)
Human insulin
A polypeptide hormone that regulates fuel metabolism and whose administration is required for survival by certain types of diabetes.
Human growth hormone
Somatotropin-induces the proliferation of muscle, bone, and cartilage and is used to stimulate growth in children of abnormally short stature
Erythropoietin
A protein growth factor secreted by the kidney that stimulates the production of RBCs & is used in the treatment of anemia arising from kidney disease
Colony-stimulating factors
Stimulate the production & activation of the white blood cells and are used clinically to counter the white cell-killing effects of chemotherapy and to facilitate bone marrow transplantation
Tissue-type plasminogen activator
(tPA) which is used to promote the dissolution of the blood clots responsible for heart attacks and stroke
Hepatitis B
Synthetic vaccines consisting of harmless but immunogenic components of pathogens
Site-directed mutagenesis
An oligonucleotide containing a short gene segment with the desired altered base sequence corresponding to the new amino acid sequence is used as a primer in the DNA polymerase I mediated replication of the gene of interest. Such a primer can be made to hybridize to he corresponding wild-type sequence if there are only a few mismatched base pairs, and its extension, by DNA polymerase I, yields the desired altered gene
Monoclonal antibodies
A single species of antibody produced by a clone of an antibody-producing cell; can be targeted against specific proteins and hence are used as antitumor agents
Trastuzumab
Monoclonal antibody (trade name Herceptin), which binds specifically to the growth factor receptor HER2 that is overexpressed in ~25% of breast cancers, blocks HER2’s growth-signaling activity, thereby causing the tumor to stop growing or even regress
Cassette mutagenesis
Complementary oligonucleotides containing the mutation (s) of interest are chemically synthesized and annealed to create a duplex “cassette”. The cassette is then ligated into the target gene, which must therefore contain an appropriately placed unique restriction site (which can be introduced through site-directed mutagenesis; the cassette must, of course, have the corresponding sticky ends)
Allotransplantation
Transplantation between humans of organs such as hearts, lungs, livers, and kidneys
Xenotransplantation
Process in which transplant is procured from animal to human (Ex. pig organ into human), which is triggered by the foreign antigens lining the blood vessels of the xenograft (the complement system constitutes the body’s first line of immunological defenses). The hyper-acute rejection occurs because the porcine tissue lacks the human proteins that inhibit the human complement system.
Severe combined immunodeficiency disease (SCID)
Any of several genetic diseases that so impair the immune system that a victim must be kept in a sterile environment in order to survive.
Liposomes
Membrane enveloped vesicles that are designed to be taken up by specific cells
Cystic Fibrosis
One of the most common genetic diseases, is caused by a defect in a protein involved in the secretion of chloride ion in the lungs and other tissues. This causes the secretion of abnormally thick mucus, which results in recurrent and often damaging lung infections leading to early death.
SCID-X1
Which is caused by a mutation in the gene encoding the yc cytokine receptor, a receptor for certain protein growth, and survival of the white blood cells known as T cells
-The viral vector containing the yc cytokine receptor gene integrates into the genome at random, the location of the transgene may affect the expression of other genes, triggering cancer
Tay Sachs disease
Caused by the absence of the lysosomal enzyme hexosaminidase A and results in progressive neuronal dysfunction that is invariably fatal by around age 3.
Lysis
Breaking open of a cell (Context: If the protein of interest is located in the cytosol of the cell, its liberation requires only the breaking open of the cell)-Osmotic lysis, the cells are suspended in a hypotonic solution (a solution in which the total molar concentration of solutes is less than that inside the cell in its normal physiological state)
Differential centrifugation
A process in which the cell lysate is centrifuged at a speed that removes only the cell components denser than the desired organelle followed by centrifugation at a speed that spins down the component of interest.
Proteases
Enzymes that catalyze the hydrolytic cleavage of peptide bonds. Care must be taken that the protein is not damaged by these enzymes.
-Many proteins are denatured by contact with the air-water interface and, at low concentrations, a significant fraction of the protein present may be lost by adsorption to surfaces
Coupled enzymatic reaction
The product of the enzyme being assayed is converted, by an added enzyme, to an observable substance.
Antibodies
Proteins that are produced by an animal’s immune system in response to the introduction of a foreign protein and that specifically bind to the foreign protein
Radioimmunoassay
A protein can be indirectly detected by determining the degree with which it competes with a radioactively labeled standard for binding to the antibody
An enzyme-linked immunosorbent assay (ELISA)
- An antibody against the protein of interest is immobilized on an inert solid such as polystyrene
- The solution being assayed for the protein is applied to the antibody-coated surface under conditions in which the antibody binds the protein. The unbound protein is then washed away.
- The resulting protein-antibody complex is further reacted with a second protein-specific antibody to which an easily assayed enzyme has been covalently linked.
- After washing away any unbound antibody-linked enzyme, the enzyme in the immobilized antibody-protein-antibody-enzyme in the immobilized antibody-protein-antibody-enzyme complex is assayed, thereby indicating the amount of the protein present.
