Antibodies

Antibodies (Ab)
Glycoproteins that bind antigens with high specificity and affinity.
(specific – react with only one antigen )
• Produced in B cells in response to an antigen :
Activated B cells  Plasma cells  secrete antibodies
• Found in plasma.Originally identified in the serum.
• Sometimes are called gamma globulins or immunoglobulins (Ig).
• Five classes: IgG, IgM, IgA, IgE & IgD
 Basic Structure
4 polypeptide chain unit (Y shaped)
• 2 heavy chains
(five kinds:  Disulfide bond

2 light chains
(two types: ,
Carbohydrate

• Disulfide bonds CL
– Inter-chain VL

– Intra-chain CH2 CH3

CH1
Hinge Region
VH
 Basic Structure
• Variable & Constant Regions
– VL & VH: Ag binding site
– CL & CH : determine the fate of Ag
• Hinge Region: rich in proline
easy to bend and be hydrolyzed by
some enzymes Disulfide bond

• Structure of the V Region

– Hypervariable (HVR) or
Carbohydrate
complimentarity determining
regions (CDR)
– Framework regions
CL
VL

CH2 CH3
CH1
Hinge Region
VH
Antibody – another view
- variable regions of
the light chain (grey)
and the heavy chain
(yellow) form the
antigen binding site

- light chain constant

region is blue while
heavy chain constant
region is red. The
two chains are joined
by carbohydrate
(purple).
 Basic Structure
• Domains
– VL & CL Disulfide bond

– VH & CH1, CH2 ,CH3

(or CH4) Carbohydrate

• Functional properties:
– VL , VH :Bind to Ag CL
– CL , CH1 : genetic marker VL
– CH2 : complement binding site;
placental transfer (IgG) CH2 CH3
CH1
– CH3 / CH4 : binding to FcR Hinge Region
VH
complement binding site(IgM)
 Valence and avidity
• Valence: the maximum number of antigenic determinants
with which antibody can react. e.g. IgG: 2, Fab:1
• Avidity : the firmness of association between a
multideterminant antigen and antibodies produced against it.
Disulfide bond

Having multiple binding sites for Carbohydrate

antigen means multi-valence, and
can increase its avidity to antigens CL
on particles such as bacteria or virus.VL
CH1 CH2 CH3
Hinge Region
VH
 Hydrolytic Fragments: Pepsin
Papain

Fc
Fc Peptides

Fab F(ab’)2
papain
• 1IgG 2Fab + 1Fc pepsin
• Fab(fragment antigen binding) • 1IgG 1F(ab’)2 + pFc’
valence = 1 • F(ab’)2: valence=2
• Fc(fragment crystallizable) • pFc’(no biological activities)
– Effector functions
 Human Ab Classes
• IgG - Gamma ( heavy chains
4 subclasses (IgG1, IgG2, IgG3, IgG4)
have slightly different sequences in H-chain and
corresponding differences in functional activities
• IgM - Mu ()heavy chains
• IgA - Alpha ( heavy chains
2 subclasses (IgA1, IgA2)
• IgD - Delta ( heavy chains
• IgE - Epsilon ( heavy chains
 IgG
• Structure
– Monomer

IgG1, IgG2 and IgG4 IgG3

 IgG
• Structure
• Properties
– Found in vascular and extravascular spaces, secretions
– Major serum Ig: 70-75%, provide immunity to most
blood borne infectious agents
– Major antibody of the secondary immune response
– the only Ab class that can cross placenta to provide
passive humoral immunity to fetus
• Structure
IgA
– Serum
• monomer
– Secretions (secretory IgA , sIgA)
• Dimer
• J chain (joining chain)
• SP (secretory piece)
(involved in the transepithelial transport of exocrine IgA
and stabilizes IgA against proteolytic degradation)

Secretory Piece J Chain

 IgA
• Structure
• Properties
– Found mainly in serum, external secretions
(such as: colostrum, milk, saliva)
– 2nd highest serum Ig
– Major secretory Ig
– A first line of defence against microbial invaders
at mucosal surfaces
 IgM
J Chain

• Structure
– Pentamer
– Extra domain (CH4) CH4
– J chain (joining chain)

Tail
Piece
 IgM
• Structure
• Properties
– Found primarily in plasma, the surface of B cell
– 3rd highest serum Ig, <10% of serum Ig
– First Ab produced in an immune responce
– B cell surface Ig: serves as a membrane receptor
B Cell Antigen Receptor (BcR)

Ig- Ig- Ig- Ig-

 IgD
• Structure
– Monomer
– Tail piece
• Properties
– 4th highest serum Ig
Tail Piece
– B cell surface Ig:
serves as an antigen receptor
 IgE
• Structure
– Monomer
– Extra domain (CH4)
• Properties
– Least common serum Ig
– Associated with acute CH4
inflammation, parasitic
infections and allergic
reactions(such as asthma)
 Biological properties of Antibody

• Combine with antigen:

Agglutinating bacteria, neutralizing toxins,
inactivating viruses.
• Fixing and activating complement: IgG and IgM
– Lead to complement mediated lysis of the microbes or
the target cells; also lead to attraction of immune cells,
and to opsonization and phagocytosis of the target cells.
• Placental transfer: IgG4
 Biological properties of Antibody(continued)
• Binding to Fc receptor:
– ADCC(antibody-dependent cellular cytotoxicity) ：
NK cells can kill Ab coated target cells directly.
– Opsonization ：
Binding to FcR of phagocytes can enhance phagocytosis.
– Causes type I allergy ：
Binding IgE Fc to FcR of mast cells/ basophils, can trigger
degranulation and enhance the acute inflammatory response.
Over-stimulation of mast cells/ basophils by this mechanism
leads to type I allergy.
 Monoclonal antibodies (mAb)

• Polyclonal antibodies
• Monoclonal antibodies:
– monospeific
– are standard research reagents and have
siginificant clinical utility.
• Immune animals or humans
• Fusion of an immortal cell with a specific predetermined Ab-producing cell
• The hybridoma cell is immortal and synthesizes homogeneous, specific mAb
which can be made in large quantities