1/2/2024

ANTIBODIES
Tran Thi Thu Phuong, PhD, PharmD

Email: tran-thi-thuphuong@usth.edu.vn

Contents
1. General definitions

2. Basic structure of antibodies

3. Antibody classes

4. Antigenic determinants on antibodies

5. Clonal selection theory

6. Monoclonal and polyclonal antibodies

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Definition

Antibodies (Abs) are antigen (Ag) binding proteins present on the B-cell
membrane and secreted by plasma cells.

 Abs are gamma globulin (also called immunoglobulin - Ig).

Abs are present on the B-cell membrane or secreted by plasma
cells.
 Have ability to bind specifically to antigen.

Abs are in the gamma globulin region after serum electrophoresis

Before react with antigen

After reacted with antigen

Kuby Immunology 5th Ed ©Freeman and Company 2003

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Abs are present on the B-cell membrane or secreted by plasma cells

Plasma cell
B cell

Antigen
Antibody

Ab can bind specifically to Ag

Specific
binding as
Abs fit to Ag in dimensional three “lock and key”
conformations of antigenic determinant
(epitope), as such being called specific
binding.

 Basic structure of Abs

 Common structure of Abs

 4 polypeptide chains:
 2 heavy chains (H)
 2 light chains (L)
 2 H chains are identical and 2 L
chains are identical as pairs.
 Chains are linked by disulfide
bonds.

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 Basic structure of Abs

Super-variable regions
 On each chain has constant region (C) and (Ag binding sites)
variable region (V)
 At the N terminus has super-variable regions or
antigen binding regions

 Chains folded to form domains

 Heavy chains has hinge regions

Hinge region
provide the
flexibility to
Ab molecule to
capture Ag.

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 Basic structure of Abs

 Common structure of Abs

Kuby Immunology 5th Ed ©Freeman and Company 2003

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The super-variable region has 3D structure that fit to the 3D structure of epitope as “lock and key”

Janeway’s Immunology, 9/e.

(®Garland Science 2017)

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 Abs classification

 Light Chains: two types, kappa () and lambda ()

 Heavy Chains: five types, define five classes of Abs:
 gamma () chain for IgG
 muy () chain for IgM
 alpha () chain for IgA
 epsilon () chain for IgE
 delta () chain for IgD

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Type of Abs defined by 5 types of H chains

                  
         

IgG IgG IgM IgM IgD IgD IgA IgA IgE IgE
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Features of the major

isotypes (classes) of
antibodies

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 IgG

 Mol. Wt. 150,000

 ~70-75% of serum immunoglobulin.
 4 Sub-classes (IgG1, IgG2, IgG3, IgG4)
 The major antibody of the secondary immune response
 Only Ab class that can pass placenta
 Change in affinity with time

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 IgG
IgG fragments after enzyme treatment

 Fab (fragment of antigen

binding)
 Contain Ag binding sites
 Have specificity of the Ab
 Fc (fragment cristalizable)
 C’ binding site
 Fc receptor site
 Fab’2

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 IgM

 10 Heavy and 10 Light chains

 10 Fab and 5 Fc fragments
 Mol. Wt. ~900,000
 <10% of serum immunoglobulin.
 Single J Chain (15 kDA)
 The predominant "early" antibody
 Most primitive immunoglobulin
 No change in affinity with time

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 IgA
 Two forms
 Serum IgA, monomeric
 ~15-20% of serum
immunoglobulins
 2 Heavy and 2 Light chains
 2 Subclasses (IgA1 and IgA2)
 Found in serum

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 IgA

 4 heavy chains and 4 light chains (dimeric)

 J-Chain and secretory component
 The major immunoglobulin of secretions
 Not found in serum

Secretory component is produced by epithelial cell,

involves in transportation of IgA and also protect Ab
against proteolytic enzymes

Kuby Immunology 5th Ed ©Freeman and Company 2003

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Transportation and secretion of IgA dimer

Kuby Immunology 5th Ed ©Freeman and Company 2003

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 IgD

 Very low quantity in serum (30µg/ml).

 On surface of mature B cell, serves as B cell

receptor for Ag.

 May play role in antigen-stimulated lymphocyte

differentiation Mature B cell

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 IgE

 Trace serum protein

 Most bind on surface of basophil and
mast cells, associated with allergy or
anaphylactic hypersensitivity
 May play role in immunity to helminthic
parasites

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Janeway’s Immunology, 9/e.

(®Garland Science 2017)

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Mast cell degranulation

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IgE and eosinophil kill parasite

Abbas A. K, Lichtman A. H, and Pillai S.

Basic Immunology 6th Ed © Saunders 2020

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Kuby Immunology 5th Ed

©Freeman and Company 2003

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Distribution of Ab classes

IgG: blood and extracellular spaces

IgM: Blood
IgA: Secretion fluid
IgE: Under skin and mucous
membrane
IgD: B cell surface

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Notes

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Antigenic determinants on Ab

 Once Ab being transferred from one body to the other, it becomes Ag the
new body.

 The differences among Ab molecules The antigenic determinants, or

epitopes, on immunoglobulin molecules fall into three major categories:
isotypic, allotypic, and idiotypic determinants, which are located in
characteristic portions of the molecule.

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Clonal selection theory

 In fetus, many B cells are produced, each can react with one epitope which
may exist in nature.
 B cells which react with epitope belonging to self components are removed or
suppressed.
 Each mature cell may have several identical copies which form a B cell clone.

 Once an Ag enter the body, B cells will come to contact the Ag. THE ONE
which has receptor (Sig) can bind to epitopes on Ag is the SPECIFIC one
being “SELECTED” for proliferation to produce mass number of SPECIFIC
cells producing Abs specific to the uncounted Ag.

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MacFarlane Burnet’s theory (1957), Nobel prize 1960

On the selection of specific B cell for Ab production

Kuby Immunology 5th Ed

©Freeman and Company 2003

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Sir Frank Macfarlane Burnet

(1899-1985)
“One can think of the middle of the twentieth century as the end of one of
the most important social revolutions in history, the virtual elimination of
the infectious disease as a significant factor in social life”
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B cell selection
 Each B cell has (pre-made) receptors that can bind
only to ONE epitope Binding to epitope is signal to
activate the cell

 Progeny cell have the same specificity (BCR and

secrete Ab) as mother cell

 Selection of one cell to activate it to proliferate to

become a clone (many cells) that increase the
frequency of Ag- specific cells encounter existing
Ag in the body

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Abbas A. K and Lichtman A. H

Basic Immunology 3rd Ed © Saunders 2011

Burnet’s Clonal
selection theory
turned to be
applicable to both
B cells and T cell

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Polyclonal and monoclonal Abs

 Polyclonal Ab is mixture of Abs produced by many B cell clones; each clone

is specific for one epitope of an Ag.

 Monoclonal antibody (mAb) is Ab produced by a single B-cell

clone and specific to one epitope only.

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César Milstein and Georges Köhler

Nobel prize, 1984
Principle of mAb production

Fuse a Ab producing B cell with a

myeloma cell (cancer cell) to form a
hybridoma cell that immortalized
(inherited from cancer cell) and
producing Ab (inherited from B cell). The
cell proliferate to form a clone producing
Ab to a single epitope.
Kuby Immunology 5th Ed ©Freeman and Company
2003
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pAb and mAb

Kuby Immunology 5th Ed

©Freeman and Company 2003

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Kuby Immunology 5th Ed

©Freeman and Company 2003

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Applications of mAb

Diagnosis
 No cross-reactivity
 Specific diagnose

Treatment
 Targeted therapy
 Reduce side effects

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