ANTIBODY

ANTIBODY
Antibodies- Immunoglobulins
 Antibodies are defined as specialised proteins

that are formed in response to an antigen.
 They react with that antigen (or a closely

related antigen) specifically and in some
observable manner.
 Formed in serum, body secretions and some are

fixed to body cells or tissues
Properties of Antibodies
 Produced by vertebrates only
 Chemical nature of antibodies is globulin and

hence named as immunoglobulins.
 Immunoglobulins are glycoproteins and

constitute about 20 – 25% of the total serum
proteins.
 Serum globulins can be water soluble

(pseudoglobulins) and water insoluble
(euglobulins)
 Most antibodies are euglobulins.
 Serum antibodies mostly migrate

electrophoretically in the Gamma region - called as
Gamma Globulins.
 Most antibodies belong to the 7S class

(MW:150,000-180,000)
 Heavier antibodies belong to the 19 S class

(MW:900,000) and are designated as (M)
Macroglobulins
 Immunoglobulins are synthesized by plasma cells.
 Immunoglobulin includes, besides antibody
globulins, the abnormal myeloma proteins, the
cryoglobulin and the macroglobulin.
 Thus all immunoglobulins may not be antibodies

but all antibodies are immunoglobulins.
Structure of immunoglobulin
 Y shaped 4 polypeptide
chain molecule
 2 Light (L) and 2 Heavy

(H) chains
2 H chains are held

together by 1-5
disulphide bonds
 L chain is attached to H
chain by 1 disulphide bond
Structure of Immunoglobulin
 L chains have a MW of 25,000 daltons
 H chains have a MW of 50,000 daltons
 H chains are structurally and antigenically
distinct for each class of Immunoglobulin.
 H chains are named as Alpha, Delta, Gamma,

Epsilon, Mu, in the Immunoglobulins IgA, IgD, IgG,
IgE & IgM respectively.
 L chains are of 2 types – Kappa & Lambda
 Named after Korngold & Lapari who described

them
 One Ig molecule can have 2 Kappa or 2

Lamdba chains but never both
 K : L = 2:1
CLASSIFICATION
 Five different classes namely IgG, IgM, IgA,
IgD, IgE are designated depending on presence of
heavy chain, gamma, mu, alpha and delta and
epsilon respectively.
 Structure of Immunoglobulin G has been studied
extensively
Aminoacid sequences
 Each light chain contains @ 214 (210 -230)
aminoacids whereas a heavy chain has 420-460
aminoacids.
 In the L chain, of the 214 aminoacid residues,

107 constitute the carboxy terminal half and occur
in constant sequence – those determining the
kappa or lambda specificities .
 This region is called the Constant region (C).

 The amino acid sequence in the amino terminal
half of the L chain is highly variable .
 This is called Variable region (V).
 H chain also has Constant and Variable regions.
 Variable (V) regions are present at amino

terminus (NH2) and Constant ( C) region at
carboxy terminus (COOH).
 In the L chain the two regions are of equal
length
 In the H chains the variable region

constitutes only the fifth of the chain.
 Antigen combining site is at its
amino terminus which
contains both H and L chains.

 The infinite range of antibody specificity of
the Immunoglobulins depends on the variability
of the aminoacid sequences at the variable
regions of H and L chains which form the antigen
binding sites
 The amino acid sequence of the variable
regions of the H and L chains are not uniformly
variable.
 They consist of invariable and some highly

variable zones
 The highly variable zones are 3 in L chains and
4 in the H chains
 They are called as Hypervariable regions or Hot
spots.
 They are involved with formation of Antigen

binding sites.
 The sites on the highly variable regions make

actual
contact with the epitope and are called

Complementarity determining regions (CDR).
Papain digestion
Pepsin digestion
 Each Ig molecule has internal
disulphide links in addition to
inter chain disulphide links.
 The inter chain disulphide

links form loops in the peptide
chain.
Each loop is compactly folded to form a

globular domain.
 Each domain has a specific function
 Variable region domains VL & VH form a specific

antigen binding site.
 CH2 region in IgG binds to C1q fragment in

classical complement activation.
 CH3 mediates
monocyte adherance.
 Fc fragment is composed of the Carboxy terminal
portion of the H chain. No antigen combining
activity
 Determines the biological properties of the

immunoglobulin molecule –
1)Complement fixation
2) Placental transfer
3)Skin fixation
4)Catabolic rate
 Portion of H chains in the Fab fragment is Fd
piece
 H chain has a Carbohydrate moiety which is
distinct for each Ig class
 This is the region at which the arms of the

antibody molecule forms a Y.
 called the Hinge region - there is more

flexibility in the molecule at this point.
 More exposed to enzymes and chemicals.
Papain acts here to produce 1 Fc and 2 Fab

fragments
Immunoglobulin G
 Major serum immunoglobulin
(about 80% of the total amount).
 Normal serum Conc. is
about 8-16mg/ml
 MW :150,000
 Half life is about 23 days (longest)

Immunoglobulin G
Equal intravascular and
extravascular distribution
 Late antibody - It appears

late but persists for
longer time.
 Appears after the initial

immune response which is
IgM in nature.
Immunoglobulin G
 IgG binds to micro
organisms and enhances
phagocytosis.
 Four subclasses of IgG:

IgG1- 65%, IgG2-,23%
IgG3-8%,IgG4-4%
 These can be identified

with specific antisera.
Immunoglobulin G
 It is the only immunoglobulin to cross the
placental barrier and provides natural passive
immunity to new born.
 Immunological reactions – Precipitation,
Complement fixation, Toxin neutralisation.

Immunoglobulin G
 General purpose antibody – protects blood and
tissues
 Unique catabolism
 Passively given IgG suppresses the

homologous IgG by a feed back process
Immunoglobulin D
• IgD resembles IgG structurally.
• Concentration of 3mg /100 ml

in serum.
• Half life is about 3 days.
• IgD & IgM it are present on the surface of

unstimulated B lymphocytes in blood and acts as
recognition receptors for antigens.
• 2 subclasses are there IgD1 and IgD2

Immunoglobulin M
 It constitutes about 5-8 % of total
serum immunoglobulins.
 The normal level in serum is 0.5-
2mg/ml.
 It is heavy molecule (19S) with a MW: 900,000 to
1,000,000 - also called “millionaire molecule”.

Immunoglobulin M
• Pentamer consisting of 5 immunoglobulin (4
peptide) subunits
• One molecule of J chain joins the Fc region of the
basic subunits
• Each H chain of IgM has four CH domains rather
than three as seen in H chain of IgG molecule.
• Half life is about 5days- Short lived.
• Presence in serum indicates
recent infection
Immunoglobulin M
• Early in response to infection before IgG
• Mainly distributed intravascularly (80%)
• Earliest synthesised immunoglobulin by foetus

in about 20 weeks of age.
• Oldest Immunoglobulin class
• Not transported across the placenta

• Presence of IgM antibody in serum of newborn
indicates congenital infection
Immunoglobulin M
• very effective antibody in agglutination,
complement fixation, opsonisation
and immune haemolysis.
It is more efficient than IgG in these reactions.
• Less efficient in toxin & virus neutralisation.
• The greater efficacy is due to the binding of

complement to several Fc regions of pentameric
IgM simultaneously, thus initiating complement
cascade and target lysis with a single molecule.
Immunoglobulin M
• Unique structure to provide protection against
microorganisms and other large antigens with
repeating antigenic determinants- Valency
• IgM provides protection against blood invasion by
microorganisms. Its deficiency is often associated
with septicaemia
Immunoglobulin M
 Iso hemagglutinins (anti A, anti B) and many
other natural antibodies are IgM.
 Typhoid ‘O’ antibodies
 Reagin antibodies in syphilis
 Monomeric IgM is the surface antibody

receptor on the
B cell surface for antigen recognition.

Immunoglobulin A
• IgA is the second

major serum
immunoglobulin (about
10- 13%) .
• The normal serum

concentration is (0.6-
4.2mg/ml).
Immunoglobulin A
• Half life is about 6-8 days
• IgA occurs in two forms, serum IgA and secretory

IgA
• Serum IgA is a monomeric molecule
• IgA found on mucosal surfaces and in secretions

are in dimeric forms
Immunoglobulin A
• It is a dimer formed by
two monomer units
joined together by a
glycoprotein named a
the J chain
• The J chain joins the

two monomer units at
their carboxyterminals.
Immunoglobulin A
• Secretory IgA and J chain are produced by

the plasma cells situated near the mucosal or
glandular epithelium.
• J chains are also present in other polymeric

immunoglobulins such as IgM
Immunoglobulin A
• Secretory IgA contains

another glycine rich
polypeptide called the
secretory piece or secretory
component.
• This is the principle

immunoglobulin present in
secretions such as milk, saliva,
tears, sweat, nasal fluids,
colostrum, etc.
Immunoglobulin A
• IgA does not fix complement but can activate

alternative pathway.
• IgA is mainly synthesised locally by plasma cells

and little is derived from serum.
• Two subclasses of IgA (IgA1and IgA2) are

known.
• IgA2 is predominant (60%) in the secretions

but constitutes a minor part of serum IgA
Immunoglobulin E
• IgE is mainly produced in the linings of

respiratory and intestinal tracts. Serum
contains only traces.
• It is also referred to as Reagin
• MW – 160,000
• Half life is 2-3 days.
• It resembles IgG in structure

Immunoglobulin E
• It has affinity for surface of tissue

cells, particularly mast cells of the
same species.
• IgE mediates type 1 hypersensitivity

reaction- anaphylaxis
• This is responsible for asthma, hay

fever, eczema and Prausnitz-Kustner
reaction
Immunoglobulin E
• High level of IgE in serum is also

seen in children with a high load
of intestinal parasitism
• It cannot cross the placental

barrier or fix complement.
Role of Different Ig classes
 IgG- protects the body fluids
 IgA- Protects the body surfaces
 IgM- Protects the blood stream
IgE - mediates reaginic

Hypersensitivity
 IgD recognition molecule on the surface of B

lymphocytes
Abnormal Immunoglobulins
• Structurally similar proteins apart from Ab are
seen in certain pathologic conditions
• Bence Jones proteins – Multiple myeloma
• Waldenstrom’s macroglobulinemia
• Heavy chain disease
• Cryoglobulinemia
• Idiotypes
Idiotypes are antibodies that recognize
different specific epitopes. The thing that
determines the idiotype is way at the end
of the variable region; it’s composed of a
bunch of different idiotopes (or combining
sites).
• An idiotype can be defined as the specific
combination of idiotopes present within an
antibodies complement determining
regions (CDRs). A single idiotope, is a
specific region within an antibodies Fv
region which binds to the paratope
(antigenic epitope binding site) of a
different antibody. Therefore, and idiotope
can be considered almost synonymous
with an antigenic determinant of an
antibody.
• Isotypes
Antibody isotypes are the same thing as
antibody classes. There are 5 major
isotypes: IgM, IgD, IgG, IgE, and IgA. The
difference between these isotypes lies in
the heavy chain (Mu, Delta, Gamma,
Epsilon, or Alpha).
• Allotypes
Allotypes represent the genetically
determined differences in antibodies
between people.
Thank you

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ANTIBODY

 Antibodies are defined as specialised proteins

 They react with that antigen (or a closely

 Formed in serum, body secretions and some are

 Chemical nature of antibodies is globulin and

 Immunoglobulins are glycoproteins and

 Serum globulins can be water soluble

 Serum antibodies mostly migrate

 Most antibodies belong to the 7S class

 Heavier antibodies belong to the 19 S class

 Immunoglobulin includes, besides antibody

globulins, the abnormal myeloma proteins, the

cryoglobulin and the macroglobulin.

 Thus all immunoglobulins may not be antibodies

 2 Light (L) and 2 Heavy

2 H chains are held

 H chains have a MW of 50,000 daltons

 H chains are structurally and antigenically

distinct for each class of Immunoglobulin.

 H chains are named as Alpha, Delta, Gamma,

 Named after Korngold & Lapari who described

 One Ig molecule can have 2 Kappa or 2

IgD, IgE are designated depending on presence of

heavy chain, gamma, mu, alpha and delta and

 Structure of Immunoglobulin G has been studied

 In the L chain, of the 214 aminoacid residues,

 This region is called the Constant region (C).

half of the L chain is highly variable .

 This is called Variable region (V).

 H chain also has Constant and Variable regions.

 Variable (V) regions are present at amino

 In the H chains the variable region

 Antigen combining site is at its

amino terminus which

contains both H and L chains.

the Immunoglobulins depends on the variability

of the aminoacid sequences at the variable

regions of H and L chains which form the antigen

regions of the H and L chains are not uniformly

 They consist of invariable and some highly

 The highly variable zones are 3 in L chains and

 They are involved with formation of Antigen

 The sites on the highly variable regions make

contact with the epitope and are called

 The inter chain disulphide

Each loop is compactly folded to form a

 Variable region domains VL & VH form a specific

 CH2 region in IgG binds to C1q fragment in

 Determines the biological properties of the

 This is the region at which the arms of the

 called the Hinge region - there is more

 More exposed to enzymes and chemicals.

Papain acts here to produce 1 Fc and 2 Fab

(about 80% of the total amount).

 Normal serum Conc. is

 Half life is about 23 days (longest)

 Late antibody - It appears

 Appears after the initial

 Four subclasses of IgG: