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29 Cards in this Set
- Front
- Back
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difference between antiserum and mAb
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the major characteristic of antisera is the heterogeneity of the Ab preparation, and monoclonal Ab has an increased specificity
in the lab, the mAb should always be used if available |
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what is the cornerstone in the generation of monoclonal antibodies
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a fusion of B cells from the spleen of an immunized mouse with myeloma cells...you get a hybridized cell that can produce high amounts of Abs and propogate indefinitely
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preparation to use against the venom of snakes
(antiserum vs. mAb) |
polyclonal antiserum (not mAb)
It has a variety of Abs that are different to neutralize different parts of snake venom (e.g. neurotoxins, antiplatelets and procoagulants) |
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term that refers to the amount of sites available on Abs and antigens for forming complexes with each other
for agglutination to occur, there must be multiple sites for interaction on both Ab and Ag |
valence
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what does the Heidelberger-Kendall curve for immunoprecipitation tell us
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the greatest precipitation occurs when there are roughly equivalent amounts of Ag and Ab in the mixture
when excess Ag or Ab is present, precipitation decreases |
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what preparation is usually used for immunoagglutination
(antiserum vs. mAb) |
polyclonal antiserum representing a population of different antibodies
monoclonal antibodies is usually not effective - unless there are several identical epitopes on the macromolecule |
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one of the earliest and simplest methods for studying Ab-Ag reactions
involves migration of Ab and Ag towards each other in a semisolid agar gel...visual percipitate forms when the concentrations become equivilant antiserum placed in the center and each Ag is placed in surrounding wells |
Ouchterlony immunodiffusion
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two stages of immunoelectrophoresis
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1st stage: electrophoresis
2nd stage: immunoprecipitation |
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major purpose of immunoelectrophoresis in medical laboratories
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to identify monoclonal immunoglobulins
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five fractions of proteins on electrophoresis
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albumin
alpha 1 globulins alpha 2 globulins beta globulins gamma globulins |
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two types of agglutination reactions
how are they different? |
direct and indirect assays
direct assays are for poylvalent Ags only , and indirect can be used for small Ags w/ a single epitope |
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what is a hapten molecule
how are they detected? |
a small Ag w/ a single epitope
they are detected by indirect agglutination reactions |
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assay that is also called "rocket" or "Laurell rocket"
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ElectroImmunoAssay
(a.k.a. EIA) (Note: this is NOT "ELISA") Ab mixed in the gel and migrates to point of equivalency, where it percipitates in an arrowhead or "rocket" design. |
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test in which whole blood is incubated with a cocktail of Abs, if the RBC has Ags that are recognized by the Abs crosslinking w/ unwanted cells will occur
immuno-rosettes are formed What does it detect |
hemagglutination (RosetteSep)
used to detect RBC ags or Abs present in the sample against RBCs |
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test that uses 96-well microplates
Ag "sandwiched" by capture and detection mAbs color reaction used to tell results |
ELISA: Enzyme Linked ImmunoSorbent Assay
(Note: This is NOT "EIA") |
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modified version of the ELISA that uses immune cells instead of fluids
function of cells analyzed, as opposed to surface Ag |
ELISPOT
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when not to separate cells based on density
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when you are trying to obtain individual cells
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4 techniques that can be used to separate cells
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based on cell density
based on adherence positive or negative selection based on cell-surface Ags |
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cells that can be separated based on adherence
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macrophages can be separated from T and B lymphs, which can then be separated from each other
Macrophages will adhere to a petri dish while T/B cells will not. |
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process in which a marker that identifies the required subset of the cells is picked up by the separation method
a process in which a marker is used, and the cells that are left behind are the wanted ones, subtraction method what molecules are used in these processes |
positive selection
negative selection CD antigens on leukocytes |
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two techniques used to separate cells using CD antigens
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fluorescence-activated cell sorting (FACS)
magnetic separation |
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test used to determine the load of a virus in the blood
first uses cDNA from mRNA, and then amplifies the gene of interest |
PCR
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technology used to measure gene expression, always relative to a reference sample
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cDNA microarray
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Seperation media used for seperation of cells based on cell density?
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Hypaque or Iospaque - iodinated benzoic acid derivative
Ficoll - dense polymer of sucrose Percol - colloidal nontoxic silica particle |
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What is the CD molecule that allows Leucocytes and T cells to be identified and seperated out?
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Leucocytes - CD45
T-cells - CD3 |
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How does the FACS seperation test differ from the magnetic seperation test in terms of purpose?
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FACS - used to anylaze the number of different cell types that have the immunoflufecent antibody bond to them
Magnetic Separation - can be used for bulk production of cells |
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How does the FACS test work
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labeled antibodies are added to suspension that has the cells to be anylazed...the solution is sent through a lazer beam and its scattered light is anylazed
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How does the Magnetic seperation technique work?
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Can be used for positive or negative selection
Uses anti-CD antigens bound to magnetic particles...a magnet is brought next to the tube and the magnetic bead-bound cells are seperated from the mixture |
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Who is the most amazing person in the whole world
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TRACY
(Note: Tracy smells!!!!!!!!!!) |
