Antibodies and Antigens

Lec 4

Asst. Prof. Dr. Deniz Balcı

deniz.balci@emu.edu.tr
 OUTLINE
• Antibody Structure, pg; 88-95
• Antigen Structure, pg;99-101
• Structure-function Relationships In Antibody
Molecules, pg;102-105
 Antigen

Any substance which

when introduced into
body, is capable of
provoking an immune
response (antibodies
production)
1) IMMUNOGENS
2) TOLEROGENS
3) HAPTENS
4) MITOGENS
3
 IMMUNOGEN
Immunogen is a substance that induces a specific immune
response and the phenomenon is known as immunogenicity.

• Proteins, polysaccharides
coats, capsules, cell walls, flagella, fimbrae, and toxins of
bacteria, viruses, and other microorganisms.

• Lipids and nucleic acids are antigenic only when combined

with proteins and polysaccharides.

• Non-microbial exogenous (non-self) antigens

pollen, egg white, and proteins from transplanted tissues and
organs or on the surface of transfused blood cells.
FEATURES of A GOOD IMMUNOGEN
Parameter Immunogenicity
Size Large > Small
Chemical Proteins>Carbohydrates>Lipids
Composition
Similarity to self- Multiple differences > Few
antigens differences
Dosage and route Subcutaneous > Oral
of administration
Use of adjuvants Increase titer and affinity
Genetic Major Histocompatibility
composition of Complex (HLA)
recipient
 Antigens Characteristics

• Foreignness: Molecules recognized as “self” are

not immunogenic
• Molecular Size: Small foreign molecule with
molecular weight below 10,000 D (hapten)
weakly immunogenic & must be coupled to
carrier molecule to be antigenic
– Once antibodies are formed they recognize hapten

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 HAPTEN

Cortisol, Estradiol, Testosterone, Thyroid hormones, Penicillin

 Antigen Characteristics
• Chemical–Structural Complexity: Amino acid

homopolymers less immunogenic than

heteropolymers (containing two or three
different amino acids)

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 Antigens Characteristics
• Antigenic Determinants (Epitopes) :Small chemical groups on
antigen molecule that can elicit immunological response & react
with antibody

• Dosage , Route & Timing of Antigen administration: These

factors affect immunogenicity

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 Epitope

• Small part of antigen that

interacts with an antibody
• Any given antigen may
have several epitopes
• Each epitope is
recognized by a different
antibody
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 TOLEROGEN
Tolerogen is an antigen that invokes a specific
immune non-responsiveness due to its
molecular form.
If its molecular form is changed
a tolerogen can become an immunogen.

•Allergies
•Autoimmunity
•Transplants
 ALLERGEN

An allergen is a substance that causes the

allergic reaction. The (detrimental) reaction
may result after exposure via ingestion,
inhalation, injection or contact with skin.
 AUTOANTIGEN
An autoantigen is usually a normal protein or complex
of proteins (and sometimes DNA or RNA) that is
recognized by the immune system of patients suffering
from a specific autoimmune disease.

These antigens should under normal conditions not be

the target of the immune system, but due to mainly
genetic and environmental factors the normal
immunological tolerance for such an antigen has been
lost in these patients.
Antigen Recognizing Molecules
SUBTYPES of ANTIGENS

• T Cell Dependent
• T Cell Independent
 Types of Antigens
T-independent
The antigens that are capable of
Ø Polysaccharides, lipids inducing antibody production
l Properties without T cells
l Polymeric structure
l Polyclonal B cell activation
l Yes -Type 1 (TI-1)

l No -Type 2 (TI-2)

l Resistance to degradation

l Examples
l Pneumococcal polysaccharide, lipopolysaccharide, Flagella
 Types of Antigens
T-dependent
Ø Proteins The antigens that require T
lymphocytes to induce immune
l Structure response and antibody production
few copies of many
different antigenic Hence T-dependent antigens do
determinants not directly stimulate the
production of antibody without the
l Majority of antigens
help of T cells.
l Examples
l Microbial proteins
l Non-self or
Altered-self
proteins
 What Does the αβ T Cell Receptor
(TCR) Recognize?
• Only fragments of proteins (peptides) associated
with MHC molecules on surface of cells

• Helper T cells (Th) recognize peptide associated with

MHC class II molecules
• Cytotoxic T cells (Tc) recognize peptide associated
with MHC class I molecules
 Antigenic Determinants
Recognized by T cells
• Composition
– Proteins (some lipids)
– Sequence determinants
• Processed (linear epitope)
• MHC presentation (lipid presentation by MHC-like
CD1)
• Size
– 8-15 residues
• Number
– Limited to those that can bind to MHC
 What Does The B Cell
Immunoglobulin (Ig) Receptor
Recognize?
• Proteins (conformational determinants,
denatured or proteolyzed determinants)
• Nucleic acids
• Polysaccharides
• Some lipids
• Small chemicals (haptens)
 Antigenic Determinants
Recognized by B cells and Ab
• Composition
– Proteins, polysaccharides, nucleic acids
– Sequence (linear) determinants
– Conformational determinants
• Size
– 4-8 residues
• Number
– Limited (immunodominant epitopes)
– Located on the external surfaces of the Ag
 Antigenic determinant or epitope:
The structure recognized by an antibody

Ag’s can bind in pockets or grooves, or on extended surfaces in the binding sites of Ab’s.
 Factors Affecting Measurement of
Ag/Ab Reactions
• Affinity
• Avidity
• Ag:Ab ratio
Ab excess Ag excess
• Physical form of Aga

Equivalence – Lattice formation

dissociation constant (Kd),
Ch. 6
 Valency and avidity of antibody antigen
interactions

interact with a single binding

site of one antibody molecule.

repeated determinants on a cell

surface are close enough, both the
antigen-binding sites of a single IgG
molecule can bind

IgM molecules have 10

identical antigen-binding sites
 CROSS REACTIVITY
• The ability of an individual Ab combining site to react with
more than one antigenic determinant.
• The ability of a population of Ab molecules to react with
more than one Ag.

Cross reactions
Anti-A
Anti-A Ab Anti-A
Ab Ab

Ag B Ag C
Ag A
Shared epitope Similar epitope
 SUPERANTIGENS
• Proteins produced by pathogens

• Not processed by antigen presenting cells

• Intact protein binds to variable region of β chain on

TCR of T cells and to MHC class II on antigen
presenting cells (APC)

• Large numbers of activated T cells release cytokines

having pathological effects “Cytokine storm”
 SUPERANTIGENS
• Definition
– Polyclonal T cell
T cell T cell
response
TCR TCR
• Examples
Ag SuperAg
– Staphlycoccal
MHC MHC
II II
enterotoxins
APC APC – Toxic shock toxin
 FUNCTION of ANTIBODIES

Antigen-binding sites of antibodies are

complementary to epitopes

Antibodies function in several ways:

- Activation of complement and inflammation
- Neutralization
- Opsonization
- Killing by oxidation
- Agglutination
- Antibody-dependent cellular cytotoxicity
(ADCC)
 Antibody Structure
All AB share the same basic structural
characteristics but display remarkable
variability in the regions that bind antigens.
Different clones of B cells produce different
antigen-binding site

AB has symmetric core structure composed of

two identical light chains and two identical
heavy chains

Both HC and LC consist of amino terminal

variable (V) regions that participate in antigen
recognition and carboxy-terminal constant (C)
regions; of the heavy chains mediate effector
functions.

Fab (fragment, antigen binding)

Fc (fragment, crystallizable).
 CLASSES of ANTIBODIES
• Gamma globulin proteins that react specifically with antigen that
stimulated their production
• 20% of plasma protein
• Five classes of antibodies:
(based on differences in heavy chains)
• IgM–first antibody produced
• IgG–most common and longest-lasting
antibody
• IgA–associated with body secretions
• IgE–involved in response to parasitic
infections and allergies
• IgD–Naive B cell Antigen receptor
Ig FAMILY

endothelial adhesion molecules

ICAM-1 and VCAM-1
 Complementarity determining
regions (CDRs)
Most of the sequence differences and
variability among different antibodies
are confined to three short stretches
in the V region of the heavy chain and
to three stretches in the V region of the
light chain. These segments of the
greatest diversity are known as
hypervariable regions.

Because these sequences form a surface

that is complementary to the three-
dimensional shape of the bound
antigen, the hypervariable regions are
also called complementarity
determining regions (CDRs)
CDR1, CDR2, and CDR3-the most
variable
CDR – COMPLEMENTARY
DETERMINING REGION
 Levels of variation in Igs
Isotypes
Antigenic (amino acid) differences
in constant regions of heavy chains
IgG and IgM-A
All present in a normal individual

Allotype
Genes coding for L and H chains are
polymorphic, and individuals can
have different alleles.

Idiotype
Antigenic determinants formed by
specific amino acids in hypervariable
region specific clone of B cells
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 CLASSES of ANTIBODIES

Antibody molecules can be divided into distinct classes and subclasses on the
basis of differences in the structure of their heavy chain C regions.
FUNCTION of HINGE REGION

Antibody molecules are flexible, permitting them to

bind to different arrays of antigens
 Membrane and Secreted forms of
Ig heavy chains
secreted form
blood and other
extracellular fluids,
hydrophilic

membrane-bound form
a hydrophobic α-helical
transmembrane region,
an intracellular
positively charged
stretch
PRODUCTION of ANTIBODIES
 Monoclonal AB’s Practical
Applications
• Identification of phenotypic markers unique to
particular cell types.

• Immunodiagnosis.

• Tumor identification.

• Therapy.
Ø TNF used to treat rheumatoid arthritis
Ø antibodies against epidermal growth factor receptors to
target cancer cells
Ø vascular endothelial growth factor (a cytokine that promotes
angiogenesis) in patients with colon cancer
 B CELL to PLASMA CELL

① increased production of the secreted form of Ig relative to the membrane IgE-2 days
form (post-transcriptional processing) IgA-3 days
IgM-4 days
② the expression of Ig heavy chain isotypes called heavy chain isotype IgG-21-28 days
switching.
Features Related to Effector Functions
 References
• Cellular and Molecular
Immunology 8th Ed.
(2015) by Abbas et al.