ANTIGENS AND

ANTIBODIES
 ANTIGENS
Chemicals that elicit an immune response
Most are proteins or large polysaccharides
from a foreign organism.

Microbes: Capsules, cell walls, toxins, viral

capsids, flagella, etc.

Nonmicrobes: Pollen, egg white , red blood cell

surface molecules, serum proteins, and surface
molecules from transplanted tissue.
 ANTIGENS
CHARACTERISTICS:

SPECIFIC REACTIVITY
Ability of the antigen to react specifically with the
antibodies or cells it provoked

IMMUNOGENICITY
Ability to provoke an immune response
immunogen allows the body to form antibodies
 ANTIGENS
Epitope:

Small part of an antigen that interacts with an

antibody.
10-12 amino acids
Any given antigen may have several epitopes.
Each epitope is recognized by a different
antibody.
EPITOPES: ANTIGEN REGIONS THAT
INTERACT WITH ANTIBODIES
 ANTIGENS
HAPTEN:
Possess reactivity but no immunogenicity
o SIMPLE OR NONPRECIPITATING
Can combine with antibody
Cannot produce precipitates

o COMPLEX OR PRECIPITATING
Can combine with antibody
Can produce precipitates
ANTIGENS
 ANTIGENS

CARRIER / SCHLEPPER MOLECULES

Larger molecules attached to haptens which

confer new antigenic specificities
FACTORS AFFECTING
IMMUNOGENICITY
FOREIGNNESS

Autoantigen: ____________________
Alloantigen: _____________________
Heteroantigen: ____________________
Heterophile antigen: __________________
FACTORS AFFECTING
IMMUNOGENICITY
FOREIGNNESS
Graft transplanted organ
Autograft: ____________________
Isograft / Syngraft: _____________________
Allograft : ____________________
Heterograft / Xenograft: __________________
FACTORS AFFECTING
IMMUNOGENICITY
SIZE

Potential antigen: ____________________

Good immunogen: _____________________
Excellent immunogen: ____________________
FACTORS AFFECTING
IMMUNOGENICITY
CHEMICAL COMPOSITION & COMPLEXITY

Proteins: ____________________
Polysaccharide: _____________________
Lipids: ____________________
Nucleic acid: ________________
FACTORS AFFECTING
IMMUNOGENICITY
ROUTE, DOSAGE AND TIMING

IV and IP routes are effective; ID offers stronger

stimulus than subcutaneous and IM route (with
exceptions)
Dose response partially dependent on nature
of immunogen processing
FACTORS AFFECTING
IMMUNOGENICITY
ADJUVANT
Substances added to vaccine and less
immunogenic substances to enhance immune
response
Stimulate B cells
Stimulate T cells
Stimulate phagocytic cells
 ANTIBODIES
Proteins that recognize and bind to a particular antigen
with very high specificity.
Made in response to exposure to the antigen.
One virus or microbe may have several antigenic
determinant sites, to which different antibodies may
bind.
Each antibody has at least two identical sites that bind
antigen: Antigen binding sites.
Belong to a group of serum proteins called
immunoglobulins
 ANTIBODIES

FUNCTIONS:
Neutralize toxic substances
Facilitate phagocytosis and kill microbes
Combine with antigens on cellular surface and
cause the destruction of these cells either
extravascularly or intravascularly
 ANTIGEN ANTIBODY
INTERACTION
PRIMARY
Non-covalent interactions
Ionic, hydrogen bonds
SECONDARY
Multivalent antigens
precipitation
agglutination
ANTIBODY STRUCTURE
ANTIBODY STRUCTURE
 ANTIBODY STRUCTURE

Basic structure four chain polypeptide unit; consists of 2 heavy and 2 light
chains held together by disulfide bonds

Heavy chains Each consists of about _____ amino acids

Determine the immunoglobulin class
Light chains or ; ratio = 65% : 35%;
Both found in all classes of immunoglobulins but only one
type present in a given molecule
Disulfide bonds Holds each light chain to a heavy chain
Link the midregion of the two heavy chains
Fab fragment Fragment antigen binding
One (1) light chain & one-half ( ) of a heavy chain; amino
terminal end
Fc fragment Fragment crystalline
Carboxy-terminal end halves of the 2 heavy chains
Complement binding site
Constant region Carboxy-terminal end of the Ig molecule where aa
sequence is the same for all chains of that type
Responsible for type & ag-ab reaction that occurs
Constant region of heavy chain differs from one ab class to
the other
Variable region Amino-terminal end of the Ig molecule where aa sequence
varies
Part responsible for the specificity of a particular Ig
Hypervariable Regions within the variable region that actually form the
region antigen binding site
Changes = immense diversity of antigen binding sites can
be created
# of binding sites = VALENCE
Hinge region Flexible portion of the heavy chain
Located between 1st & 2nd constant regions
Allows molecule to bend

Joining chain Glycoprotein that serves to link Ig monomers together

TYPES OF ANTIBODIES
1. IMMUNOGLOBULIN (IgG)

Predominant immunoglobulin in humans

Approximately 75 80% of the total serum
immunoglobulins
7S molecule with a MW approx. 150,000 Da
Made up of 1 basic structural unit known as a
monomer consisting of 2 heavy and 2 light chains
1,200 mg/dL concn in plasma & serum
TYPES OF ANTIBODIES
Subclasses:
Subclass Population No. of disulfide bonds
IgG 1
IgG 2
IgG 3
IgG 4

Functions:
Providing immunity for newborn
Fixation of complement
Neutralization of toxins & viruses
Participation in agglutination reactions
TYPES OF ANTIBODIES
2. IMMUNOGLOBULIN (IgM)
Most primitive
First to appear after a primary antigenic stimulus
Made up of 5 basic structural units in circular
arrangement
Possess J chain
Functions:
Complement fixation
Agglutination
Opsonization
Neutralization of toxins
TYPES OF ANTIBODIES
3. IMMUNOGLOBULIN (IgA)
Sublcasses: IgA1 and IgA2
Concn: 200 mg/dL
Monomer: serum
IgA2 found as a dimer in body secretions
Secretory component
TYPES OF ANTIBODIES
4. IMMUNOGLOBULIN (IgD)

Most IgD present is found on the surface of

immunocompetent but unstimulated B lymphocytes
Postulated to be an anti-idiotypic antibody
Function:
immunoregulation
TYPES OF ANTIBODIES
5. IMMUNOGLOBULIN (IgE)

Lease abundant immunoglobulin in the serum

Heat labile antibody
Function:
Mediates some types of hypersensitivity and is
generally responsible for an individuals immunity
to invading parasites
Binds strongly to a receptor on mast cells &
basophils & mediate release of histamine &
heparin
ANTIBODY FRAGMENTATION
o Proteolytic enzymes
o Peptide bond splitting enzymes
o Degrade Ig molecules into definable fragments
1. Papain
Cleaves the molecule into 2 Fab fragments
& 1 Fc fragment
Directly on the hinge region
2. Pepsin
Cleaves the molecule into 2 Fab fragments
Below hinge region
ANTIBODY DIVERSITY
THEORIES
1. Paul Ehrlich Side Chain Theory
Combination of antigen with a pre-formed B cell
receptor (antibody) triggered the cell to produce
and secrete more of those receptors

2. Template theory
Instructional hypothesis
A flexible antibody molecule is acted on by the
antigen to form a complementary binding site
ANTIBODY DIVERSITY
THEORIES
3. Clonal Selection Theory
Postulates:
a. Antibodies of all specificities are produced prior to
contact with the antigen
b. B lymphocytes participating in the immune
response have receptors on their surface
membranes that are Ig molecules of the same
specificity as that of the antibodies that will be
produced by their activated and differentiated
ontogeny
ANTIBODY DIVERSITY
CLASS SWITCHING
1. During an antibody response to a T dependent
antigen a switch occurs in the class of immunoglobulin
produced from IgM to some other class except IgD

2. During class switching, another DNA rearrangement

occurs between a switch site in the intron between the
rearranged VDJ regions and the C gene and another
switch site before one of the heavy chain constant
region genes.
ANTIBODY DIVERSITY
CLASS SWITCHING
3. The result of this recombination event is to bring the
VDJ region close to one of the other constant region
genes, thereby allowing expression of a new class of
heavy chain.

4. Since the same VDJ gene is brought near a different C

gene and since the antibody specificity is determined
by the hypervariable regions within the V region, the
antibody produced after the switch occurs will have the
same specificity as before.
MONOCLONAL ANTIBODY
PRODUCTION

1. Mouse is immunized with a certain antigen

2. Spleen cells are harvested & combined with
MYELOMA cells in the presence of polyethylene
glycol
3. Plasma cell + myeloma cell = ____________ cell
4. Cells are cultured using Hypoxanthine, Aminopterin
and Thymidine medium
5. Myeloma cells and normal B cells die
MONOCLONAL ANTIBODY
PRODUCTION
6. Hybridoma cells are diluted out and placed in
microtiter wells where they are allowed to grow
7. Desired antibody is identified
8. Plasma cell + myeloma cell = ____________ cell
9. Antibody maintained in cell culture to produce a
ready supply of monoclonal antibody that reacts with
a single epitope