K4.2 : 95% in mouse : 60% in human IMMUNOGLOBULIN CLASSES.
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Transcript of K4.2 : 95% in mouse : 60% in human IMMUNOGLOBULIN CLASSES.
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K4.2
: 95% in mouse
: 60% in human
IMMUNOGLOBULIN CLASSES
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Opsonisation
agglutination
neutralisation
antibody-dependent cell-mediated cytotoxicity
complement activation
mast cell degranulation
protection of the neonate
Different immunoglobulin and their function
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IgG
• Structure– Monomer (7S)
IgG1, IgG2 and IgG4
IgG3
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IgG
• Structure• Properties
– Major serum Ig– Major Ig in extravascular spaces– Placental transfer – Fixes complement– Binds to Fc receptors
• Phagocytes - opsonization• K cells - ADCC
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IgM
• Structure– Pentamer (19S)– Extra domain
(CH4)
– J chain CH4
J Chain
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IgM• Structure• Properties
– 3rd highest serum Ig– First Ig made by
fetus and B cells– Fixes complement
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Fixation of C1 by IgG and IgM Abs
C1r C1s
C1qC1r C1s
C1q
No activation Activation
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IgM
• Structure• Properties
– 3rd highest serum Ig– First Ig made by
fetus and B cells– Fixes complement
Tail Piece
– Agglutinating Ig– Binds to Fc receptors– B cell surface Ig
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IgA• Structure
– Serum - monomer– Secretions (sIgA)
• Dimer (11S)• J chain• Secretory
component
J Chain
Secretory Piece
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IgA
• Structure• Properties
– 2nd highest serum Ig– Major secretory Ig– Does not fix complement unless
aggregated– Binds to Fc receptors on some cells
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IgD
• Structure– Monomer– Tail piece
Tail Piece
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IgD
• Structure• Properties
– 4th highest serum Ig– B cell surface Ig– Does not bind complement
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IgE
• Structure– Monomer– Extra domain
(CH4)
CH4
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IgE
• Structure• Properties
– Least common serum Ig– Allergic reactions– Parasitic infections– Does not fix complement
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14B. IgG heavy chain variability
~200 variable region segments (V)~50 diversity regions (D)4 joining segments (J)5 constant regions (C)
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15
V D J CDNA of precursor cell (not to scale)
random gene rearrangementB-cell DNA
mRNA
transcriptionand splicing
translation
C. Developmental gene rearrangements
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19polyclonalmany antibodiesthat recognize many epitopes
monoclonalone antibodythat recognizesone epitope
various classes one class
making specificantibodies requirespurified antigen
specific antibodycan be made withimpure antigen
difficultreproducibility high reproducibility
A.
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20B. Monoclonal antibody production
B-cells do not survive in culture
B-cell fused to myeloma cells produce hybridomas which survive in culture and produce antibody
Fusion is inefficient so a selectionfor hybridomas is used.
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21immunized mouse
antibodyproducing
B-cells
mutant myeloma cellsfusion
hybridomas
cell mixture
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22
well containing selective medium
test supernatant for antibody of interestdilute
retest for antibody of interest