ALA removal is accomplished by absorption using _____coated with anti-rabbit or anti-mouse
immunoglobulin for Thymoblobulin or OKT3, respectively. Correct Ans-magnetic beads
serum preparations Correct Ans-1. Recalcificiation of plasma
2. Adsorption with lymphocytes
3. Inactivation of IgM antibodies
a. DTT treatment
b. Heat Inactivation
4. Depletion of therapeutic anti-lymphocyte antibodies from serum
Why is calcium added to plasma to convert it to serum? Correct Ans-Calcium is added to
the plasma to combine with the citrate present in the anticoagulant.
This restores a normal ionized calcium level and allows clotting to occur.
At what temperature should serum absorption with lymphocytes be performed? Correct
Ans-Typically, auto-antibodies are cold reacting and should be absorbed at 4oC while
alloantibodies
are warm reacting and should be absorbed at 37oC.
DTT treatment is more specific than heat treatment in
, CHT EXAM COMPLETE STUDY GUIDE TEST WITH
SOLUTIONS
removing IgM antibodies. However, DTT can also affect the _______ in IgG antibodies.
Correct Ans-disulfide bonds
What happens if we don't remove OKT3 prior to cytotoxicity assays? Correct Ans-OKT3 is a
monoclonal antibody directed against the CD3 molecule. Therefore, the
binding of OKT3 to any CD3+ cell will activate complement and lyse the target cell. This results
in falsely positive reactivity.
Lymphocytes have a density of_______ Correct Ans-~1.077 g/ml.
Cells with a higher density than FH Correct Ans-red cells and granulocytes
cells of a lower density than lymphocytes Correct Ans-platelets
isolation of cells from lymph nodes or spleen is normally fast and simple and leads to greater
_____ and ______ Correct Ans-cell yield
viability.
Obtaining lymphocytes from spleen will require
an additional FH step along with purification steps due to a large number of contaminating
cells,
such as _______ ,______ and _______. Correct Ans-macrophages
, CHT EXAM COMPLETE STUDY GUIDE TEST WITH
SOLUTIONS
erythrocytes
platelets.
intrinsic cell properties Correct Ans-cell size and
granularity of the cell.
One method of lymphocyte isolation (HP) utilizes an intrinsic cell property, __________, by
using a
density gradient medium. Correct Ans-cell density
extrinsic cell properties Correct Ans-cell surface marker, phagocytic capabilities and
adherence capabilities.
Immunomagnetic bead separations use the extrinisic property of __________ as the
Nylon wool isolation incorporates the extrinsic property of ___________ Correct Ans-
adherence for B cells.
Nylon wool isolation procedure: Correct Ans-Lymphocytes are obtained and added to a
column packed with nylon wool. B cells will adhere to the nylon wool at an optimal
, CHT EXAM COMPLETE STUDY GUIDE TEST WITH
SOLUTIONS
temperature of 37o C. The non-adherent T cells can be collected by washing the column
several times with warm media. The B cells can then be eluted by
plunging cold media through the column.
Dead cells in a flow cytometry assay can lead to
____________ of monoclonal antibodies and/or fluorochromes. Correct Ans-nonspecific
binding
Various cell purification methods are Correct Ans-Percoll and Thrombin: remove
granulocytes, platelets, and dead cells
Lympho-kwik: isolate specific populations (T &/or B lymphocytes, lymphocytes and
monocytes)
centrifugation slow/short: remove platelets in the supernatant
carbonyl iron: Granulocytes ingest altering density (heavier)
Dnase: remove dead cells
Lymphocyte interface is
indistinctive or thin Correct Ans-Hyperlipemic blood sample
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