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28 Cards in this Set
- Front
- Back
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Where can you find the heavy chain?
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1. heavy chain - 4 chains at base/C term and four chains in the middle of the top of the Y/N term
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obj 1 - List the protein/polypeptide components of immunoglobulin molecules
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1. heavy chain - 4 chains at base/C term and four chains in the middle of the top of the Y/N term
2. light chain - four chains on the outside of the top N term of the Y 3. constant region - 4 chains in the base of the y and four chains at the base/C term of the Y 4. variable region- four chains at top of Y near N term 5. fab - variable region + the constant region in the Y towards the N term (binds antigen) 6. fac is the constant region towards the base of the Y at the C term |
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obj 1 - How do the polypeptide components of the Ig molecule interact to form a complete Ig molecule?
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1. Ig is composed of beta sheets --> connected by loops to form --> globular domains --> link together to become globular regions
2. globular regions that link the base of the Y to the top of the Y are disulfide bonds |
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What is fab? What does is its significance?
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-Fab is what you get when you cleave Ig with papain.
-Its the N term of the Ig if you cut it at the hinge. -Since it contains the variable domain it can bind antigen |
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What is fac? What is its significance?
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-Fac is what you get when you cleave Ig with papain along with Fab
-Fab cannot bind Ig because it only contains a part of the constant region |
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ob j 2 -Daigram the regions of each Ig polypeptide chain
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1) N term - light chains on outside, heavy chains on inside, constant region at top
2) C term - constant region and heavy chain 3) hinge - connects the neck of the Y with the base of the Y |
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and indicate how each region contributes to Ig function?
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1) constant region - does not bind antigen
2) hinge - allows Ig to fit to differently spaced antigen epitopes 3) variable region- binds epitope of antigen |
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How many AAs are in each beta sheet?
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110 aas
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Is there a difference between the beta sheets of the variable region and the constant region?
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Yes ( at the N term the variable region) has different beta sheets then in the constant region closer to the C term
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How many different types of heavy chains are there? light chains?
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Light Chains - 2
Heavy Chains - 5 |
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What are the different times of light chains?
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1) lambda, kappa
2) Epsilon, gamma, mu, alpha, delta |
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obj 3- What is an epitope?
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An epitope is a peice of the antigen that is on its surface and that the Ig binds to.
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obj 3 - how does an epitope interact with the Ig?
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1. Ig binds the epitope at the N term of the variable region
2. If there is an equal amount of antigen/epitope and Ig - there will be a large complex that forms and cause inflammation |
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How do you produce monoclonal antibodies?
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1. find a plasma cell that is producing the antibody you want
2. hybridize that plasma cell with a melanoma cell 3.treat the resulting cells with a drug so that only the hybridoma cells survive 4. add antigen to the cell population 5. select for those cells that have bound the antigen |
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objective 4 - How are antibodies used in diagnosis?
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1. Eliza - measure protein
2. Flowcytometry and western blot - cell seperation and characterization 3. immunohistochemistry - localization of antigen in tissue 4. immunoprecipitation and western blot -identification and characterization of antigen |
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What are three therapuetic uses of antibodies?
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1) treat inflammatory disease with anti-cytokine
2) prevent cancer using toxin conjugated antibodies 3) passive immunization |
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Please describe how to use flow cytometry to identify cell types within a population?
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1) flourscently label cells with membrane bound antibodies
2) flow sample through a laser and detector 3) the emission spectra tells you which cells expressed the particular molecular markers |
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What do you use ELiza to measure?
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protein levels
viral levels hormone levels |
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How do you do an ELIZA?
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1) add Ig specific to antigen at the bottom of a microtiter well
2) add specific amt of antigen 3) wash off excess antigen 4) add a second antibody that is specific to that antigen 5) wash of excess 2nd Ig 6) measure the amount of the 2nd antibody 7) graph the amount of the second antibody as a function of the antigen concentration and get a standard curve |
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Name three conditions that monoclonal antibodies can be used to treat?
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1)anti-IgE for allergic asthma
2)anti TNF alpha for rheumatoid arthritis 3) anti CD 20 to treat non hodgkin lymphoma type B 3) |
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What are the different types of monocolonal antibodies?
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1) human only
2) chimeric 3) humanized 4) animal only |
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What is the difference between chimeric and humanized?
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1) humanized has only mixed dna in the variable region
2) chimeric has a human constant region but the rest is animal only |
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What are chimeric, humanized and all human Ig used to treat?
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chimeric - antiCD for non hodgkin lymphoma
humanized - antiIgE for allergic asthma all human - anti TNF alpha for rhuematoid arthritis |
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What is the difference between secreted Ig and membrane bound Ig?
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1)secreted Ig participates in the humoral response to antigen
2) mIg is the B cell receptor |
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Where can you find the light chain on Ig?
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2. light chain - four chains on the outside of the top N term of the Y
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Where can you find the variable region on Ig?
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4. variable region- four chains at top of Y near N term
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Where can you find the constant region on Ig?
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3. constant region - 4 chains in the base of the y and four chains at the base/C term of the Y
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What are some of the Ig and epitope characteristics (ie does each Ig have one or more epitopes, whats the significance, etc)?
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2. Most antigens are multivalent for epitopes meaning the have more than one. These multivalent epitopes could be unique or redundant
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