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Immunoglobulin and Monoclonal Antibodies
Immunoglobulin and Monoclonal Antibodies
Immunoglobulin and Monoclonal Antibodies
Monoclonal antibodies
Reported by
Ganesh
M.Sc.D endo student
What is immunoglobulin ?
Immunoglobulin's are glycoprotein
molecules that are produced by
plasma cells in response to an
immunogen and which function as
antibodies. The immunoglobulins
derive their name from the finding that
they migrate with globular proteins
when antibody-containing serum is
placed in an electrical field
Immunoglobulin Fragments
Structure/Function Relationships
Fab
Ag binding Papain
Valence = 1
Specificity
determined by VH
and VL
Fc ( crystallizable)
Fc
Effector functions
Fab
Immunoglobulin Fragments
Structure/Function Relationships
Ag
Binding
IgM Decreases in
a) Agammaglobulinemia
b) Lymphoproliferative disorders
(certain cases)
c) Lymphoid aplasia
d) IgG and IgA myeloma
e) Dysgammaglobulinemia
f) Chronic lymphoblastic leukemia
clinical implications of human
immunoglobulin classes
IgD Increases in
a) Chronic infections
b) IgD myelomas
clinical implications of human
immunoglobulin classes
IgE Increases in
IgE Decreases in
a) Congenital agammaglobulinemia
b) Hypogammaglobulinemia due to
faulty metabolism or synthesis of
immunoglobulins
Monoclonal antibodies are antibodies
that are identical because they were
produced by one type of immune cell, all
clones of a single parent cell
Polyclonal antibodies are antibodies
that are derived from different cell lines
Epitope, also known as antigenic
determinant, is the part of an antigen that
is recognized by the immune system
specifically by antibodies, B cells, or T
cells.
In 1975, Georges Kohler and Cesar
Milstein first fused lymphocytes to
produce a cell line which was both
immortal and a producer of specific
antibodies. The two scientists were
awarded the Nobel Prize for Medicine in
1984 for the development of this
"hybridoma." The value of hybridomas to
the field was not truly appreciated until
about 1987, when MAbs were regularly
produced in rodents for diagnostics.
Monoclonal antibodies
Antibodies produced from a single
clone of B cells.
Produced by fusing a B cell secreting
the desired antibody with a myeloma
cell capable of growing indefinitely in
tissue culture.
Monoclonal antibodies all have
identical antigen-binding sites. Thus
they all bind to the same epitope with
the same affinity. They are all of the
same antibody class (isotype).
Hybridomas Technique
B lymphocytes can mutate into tumor
cells that result in a type of cancer
termed myeloma.
- Myeloma cells become “immortal”
and will grow indefinitely in culture.
- Fusion of a single activated B cell
and a myeloma cell will create a
hybridoma that can grow indefinitely in
culture
Hybridoma Selection
The “HAT Trick”
Myeloma cells have been genetically
engineered such that they can not use
hypoxanthine, aminopterin, and
thymidine (HAT medium) as a source
for nucleic acid biosynthesis and will die
in culture.
Only B cells that have fused with the
engineered myeloma cells will survive in
culture when grown in HAT medium.
Polyclonal antibodies Monoclonal
Antibodies
Ag-binding sites: A mixture of Abs with All Abs have the same
different antigen-binding antigen binding site
sites
spleen
Each cell may have produced a different antibody
spleen
fuse
“immortalized”
+
dissociate cell
… and produce a
mono clonal population
USES
Measuring protein and drug levels in
serum
From
http://users.rcn.com/jkimball.ma.ultranet/BiologyPages/A/Affi
nityChrom.html
Tumor diagnosis
in vitro in vivo
99m
Tc-mAb
Monoclonal antibodies for cancer
treatment
Three mechanisms that could be
responsible for the cancer treatment.
1. mAbs act directly when binding to a
cancer specific antigens and induce
immunological response to cancer
cells. Such as inducing cancer cell
apoptosis, inhibiting growth, or
interfering with a key function
Monoclonal antibodies for cancer
treatment
2. mAbs can be modified for delivery
of a toxin, radioisotope, cytokine or
other active conjugates.