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32 Cards in this Set
- Front
- Back
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What are a few reasons why antibodies so promising as therapeutic agents? Down sides? |
They have extremely high specificity - can target viruses, bacteria, even cancer cells. However, they may be difficult to use in vivo - may end up places you don't want them. Also, there may be multiple targets in treatment needed to see an effect. They're also only good for extracellular treatments |
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What are some things that can be targeted with antibodies? |
toxins, bacteria, viruses, cancer cells, proteins, carbohydrates, pretty much anything. They're very specific. |
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What are monoclonal antibodies and how are they structured? What are polyclonal? |
Monoclonal - single specificity Polyclonal - multiple specificities
monoclonal antibodies have two heavy chains and two light chains. The lower part of the Y is constant and the tips of the arms are variable. The variable domain is one heavy chain and one light chain |
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What are the main genetic parts in making an antibody by a lymphocyte? |
The constant region is coded by constant-region genes. Far fewer than the variable region genes. Can genetically recombine domains for specificity - gene shuffling |
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What is a primary immune response? |
B lymphocytes are exposed to antigen and begin dividing to form lymphoblasts (clones), which go on to make plasma cells that make tons of antibodies and B memory cells that store the antibody information. Essentially, whatever has the best match replicates the fastest |
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What is a secondary immune response? |
The B memory cells quickly engage with a highly specific antibody and make tons of plasma cells to secrete even more antibody. |
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What is IgM vs IgG? Which response has which one? |
Immunoglobulin M - antibodies that are secreted immediately after exposure to antigen. Immunoglobulin G - the long term antibody response. Shown more later and especially after secondary response. |
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What is the anamnestic response? |
The fast secondary response of mostly IgG to exposure to a past antigen. Gives us rapid protection from an antigen and is what allows vaccines to work |
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What is the hybridoma technique for making monoclonal antibodies? |
Inject a mouse with an antigen. When it dies, harvest the spleen. Combine with myeloma cells to make hybridomas (immortal antigen producing cells). Dilute out to one cell per micro titer and assay for desired antibody production. Clone the cells. Keep as a cell culture or infect a mouse with it (mice produce more antibodies - better culture) |
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What are some functional differences between monoclonal and polyclonal antibodies? |
Polyclonal have multiple specificities and contains many antibodies. Can't make pure polyclonal without a very pure antigen. Can make results hard to replicate due to alterior binding possibilities. Monoclonal can be made even with impure antigen starting product, single antibody, single specificity, very accurate and easy to reproduce results consistently |
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Human monoclonal antibodies: Why and how are they made? |
Mouse MoAb's can elicit an immune response in humans and get cleared out before they can help anything. Human MoAb's are less likely to elicit a response. They're made by using transgenic mice the express chimeric or humanized antibodies. |
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How could you use E. coli to produce antibodies? |
You could take the expressing cell, reverse transcribe to cDNA, fragment the heavy and light chains of human lymphocytes, separate them out and put them into a phage library. Select for best binding, do domaine shuffling and point mutations, reselect, clone sequence into E. coli. |
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How does ELISA use antibodies in diagnostic assays? |
It links them to enzymes that cause color change (measurable) when antibody and antigen connect. Can use it with antibodies or antigen fixed |
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what are some applications for human MoAbs? |
Anti-rabies glycoprotein, autoimmune diseases (anti-antibody used), many cancers, alzheimers, etc. |
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Pro-drugs: Conjugating MoAb's - pro's and cons |
Adhering a drug to an antibody for high specificity. Would go to target then drug would be activated there with low overall body toxicity. Problem: Antibodies would end up all over the place, hard to get them in a single spot. Rapidly cleared from circulation. Only acts externally of the cells |
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What are some limitations to vaccine production? |
Not all pathogens have been cultured Culturing of pathogens might not produce enough of them and requires a lot of safety, training, and expense Attenuated strains may revert Killing or attenuation might be non-uniform Limited half life and special storage conditions. Not all things can be vaccinated against (HIV, HCV) |
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What are the main types of vaccines? |
Inactivated attenuated subunit peptide rDNA |
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Inactivated vaccine |
Killed |
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Attenuated |
Live virus but made nonpathogenic by removing virulence genes. May revert under certain conditions or combine with another cell in vivo to regain virulence |
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Subunit |
Toxin or another part of the pathogen is used. |
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Recombinant DNA technology - in what ways is it utilized to make safer vaccines? |
Can be used to isolate a protein encoding gene that is particularly immunogenic instead of using whole pathogen (foot and mouth disease and HPV) Can remove virulence gene or otherwise remove things that make the pathogen grow Can program antibodies to attack only infected cells (expressed protein) |
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What is differential fluorescence induction? |
encode a library of genes (each gene per condition of bacteria), tag all with some sort of fluorescent reporter, then filter off reporting genes using FACS and identify |
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Why would you want to isolate a viral promoter? How would you do it? |
You would fluorescently tag the promoter, put it in the pathogen, infect the host, then FACS out the fluorescent showing cells. Would tell you what cell is expressing the promoter region |
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What is a hapten? |
Immunogenic only when attached to a larger molecule that is not immunogenic by itself. However, they are antigenic (cause creation of antibodies) |
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What is a peptide vaccine? What is it composed of? |
Has a carrier protein that is not immunogenic nor antigenic and is attached to a bunch of short peptides that are also not immunogenic but ARE antigenic and create immune response (immuogenic) when attached to larger molecule |
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How could you increase the effectiveness of a peptide or sub unit vaccine? |
Link to more immunogenic carrier to boost immunity, point mutations, etc. |
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What is genetic immunization? |
Using DNA to alter the genetic code of a cell which then releases viral protein that stimulates B-Cell response and also stimulates cytotoxic T-cell response |
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What are some pro's for genetic immunization? |
no need to culture infectious agents no chance of reversion to virulence fewer chance of side effects in immunocompromised peoples cheap storage and production (just DNA no protein) Can make more complex vaccine involving several antigens/vaccines repeat immunizations possible More likely to get correct protein folding |
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What are some diseases for which vaccines have been created? |
Plague, meningitis, anthrax, cholera, typhoid fever, small pox, rabies, hepatitis A and B, chickenpox |
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What are recombinant vaccines? |
Vaccines that encode one or more pathogen neutralizing agents. Can make therapeutic vaccines that can be used after infection occurs |
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What are the six ways that antibodies protect the body? |
agglutination (bind antigens together for transport out) Opsonization (agglutination then take to macrophage) Neutralization (coat and prevent from acting/replicating) Anti-body dependent cell mediated cytotoxicity (attaches to cell and causes destruction - non specific) Inflammation (attracts phagocytic cells and other immune cells/compounds) Activation of complement - complement goes into other cell triggering lysis |
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What could cause the outbreak of a disease that a vaccine exists for? |
Herd immunity could be low, there could be changes in sensitivity of test, the virus could change, or there could be an increase in mutants. |
