ANTIBODY

ANTIBODIES

 Substances produced in
response to antigenic stimulation
that are capable of specific
interaction w/ the provoking
immunogen.
 General term is immunoglobulin.
PROPERTIES OF AN
ANTIBODY
 PROPERTIES

 CHON in nature
 High molecular weight
 Present in serum/plasma, CSF,
saliva & seminal fluid
CLASSIFICATION OF
ANTIBODY
 A. ACCORDING TO THE
TEMP. @ W/C THEY REACT
 Cold Abs – react @ 4oC to
room temp. (Anti-A & B typing
sera)
 Warm Abs – react @ 37oC.
(Anti-Rh)
 B. ACCORDING TO
OCCURRENCE
 Natural Abs – appear w/o any
apparent stimulus.
 Immune Abs – appear following
the introduction of an antigen.
 C. ACCORDING TO THE
SPECIES W/C PRODUCE
THEM
 IsoAbs – produced after the
introduction of the antigen from
the same specie.
 Heterophile Abs – produced
after the introduction of the Ag
from another specie.
 D. ACCORDING TO ITS
REACTION W/ AN ANTIGEN
1. Agglutinins – responsible for immobilization
of motile organisms & for cell clumping.
2. Agglutinoids – agglutinins that are modified
by heat in such a manner that they cannot
bring about agglutination but still are
capable of combining w/ specific
agglutinogens.
3. Hemagglutinins – cause agglutination of red
cells.
 D. ACCORDING TO ITS
REACTION W/ AN ANTIGEN
4. Precipitins – demonstrated only against
soluble antigens.
5. Lysins – cause dissolution (lysis) of
antigenic cells.
• Bacteriolysin – acts upon bacterial cells.
• Hemolysin – acts upon the red cells.
• Leukocidin – kills leukocytes w/ or w/o
lysis; produced by certain bacteria.
 D. ACCORDING TO ITS
REACTION W/ AN ANTIGEN
6. Opsonins – act upon bacterial invaders,
weakening their resistance to the
phagocytizing action of leukocyte.
7. Neutralizing (protective) Abs – render
antigenic microorganisms (usually
viruses) harmless.
8. Allergenic Abs – Abs w/c react in
allergic reactions.
 D. ACCORDING TO ITS
REACTION W/ AN ANTIGEN
9. Complement fixing Abs – detected by
the consumption of the complement
by antigen-antibody complex.
10. Blocking or inhibitory Abs – combine
w/ the antigen but are not grossly
detectable unless they are shown to
block a reaction or unless the CHON
species can be ID.
E. ACCORDING TO THE NUMBER
OF STRUCTURE UNITS

 Monomer –one structure unit

 Polymers –more than a single
monomeric unit
Dimer – 2 structure units
Trimer – 3 structure units
Pentamer – 5 structure units
 STRUCTURE OF
IMMUNOGLOBULINS
 STRUCTURE

 Each of the Ig classes has a basic

structural similarity: the molecule appears
Y-shaped.
 Ig is a symmetrical 4-polypeptide unit
consisting of 2 heavy (large) & light
(small) chains.
 Varying amounts of CHO are attached w/c
vary from 45% to 18% of total composition
of Ig molecule.
 HEAVY CHAINS

 m.w. varies from 50k-90k d.

 Consist of 440 a.a. residue w/c
determines the class of Ig.
 LIGHT CHAINS

 2 types: kappa (κ) & lambda (λ )

 For any Ig molecule only 1 type, kappa
or lambda, chain is present.
 m.w. 23k d.
 Consist of 214 a.a. residue in kappa
chain.
 Consist of 213 to 216 a.a. residue in
the lambda chain.
 STRUCTURE

 The chains are held together by

disulfide bonds.
 The number of disulfide bonds between
heavy chains varies from 1 to 14.
 2 types of disulfide bonds: interchain &
intrachain.
 Globular regions folded by disulfide
bonds is called domains.
 IMMUNOGLOBULIN
DOMAIN
1. Internal disulfide bridges from loops in
the peptide chain, forming “domains”
that have distinct biological properties.
2. Light chains have 1 variable (VL)
domain & 1 constant (CL) domain.
3. Most heavy chains have 1 variable
(VL) domain & 3 constant (CH1, CH2,
CH3) domains.
 IMMUNOGLOBULIN
DOMAIN
4. IgE light chains have an extra
constant domain.
5. Variable domains are responsible
for the formation of specific
antigen-binding sites.
6. The CH2 domain in IgG binds C1q
& controls catabolic rate.
 IMMUNOGLOBULIN
DOMAIN
7. The CH3 domain in IgG is
responsible for cytotropic
reactions involving macrophages
& monocytes; attachment site for
phagocytes, platelets,
heterologous mast cells,
cytotoxic/killer cells & B cells.
 STRUCTURE

 Each chain consists of a

constant carboxyl terminal
portion & a variable amino
terminal portion each of w/c is
genetically determined by at
least one gene.
 STRUCTURE

 Variable region of Ig molecule is

encoded by the V-D (light chains)
or V-D-J (heavy chains) genes.
 The constant region is the same for
all Ig polypeptide chains of
aparticular class or type.
ANTIBODY STRUCTURE
ENZYME HYDROLYSIS
 ENZYME HYDROLYSIS

 Treatment of Ig usually occurs near

or w/in the hinge region.
 Hinge regions allows an Fg
molecule to change its configuration
from one floating Ig molecule w/ a T
configuration to a Y shape when the
Ig molecule binds to the Ag.
 ENZYME HYDROLYSIS

 2 enzymes have been useful

in determining the functions of
specific areas:
1. Papain
2. pepsin
 1. PAPAIN

 Splits the ff. fragments:

1. 2 Fab (fragment capable of
antigen binding) – composed
of Lc and ½ of 1 Hc.
 1. PAPAIN

2. 1 Fc (fragment crystallizable
upon purification) – consists of
the remainderof the constant
region of the 2Hc linked by
disulfide bonds; carries no Ab
activity but only biological
activity such as:
 1. PAPAIN

• Complement fixation
• placental permeability/transport
• attachment to phagocytic cells
• degranulation of mast cell
• skin fixation
• histamine release
PAPAIN HYDROLYSIS
 PEPSIN

 Results in the ff. fragments:

1. F(ab)’2
2. F(c’)
PEPSIN HYDROLYSIS
IMMUNOGLOBULIN
(VARIABILITY)
HETEROGENEITY
IMMUNOGLOBULIN
HETEROGENEITY

1. Isotypic variation – refers to the

different H & L chain classes &
subclasses.
2. Allotypic variation – refers to the
genetic variation w/in species
involving different alleles @ a given
locus; occurs mostly in the
constant region.
 IMMUNOGLOBULIN
HETEROGENEITY
3. Idiotypic variation - refers to
the diversity @ the binding site
& in particular relates to the
hypervariable segments of the
Ab combining site.
 TYPES OF
IMMUNOGLOBULIN
 IgG

 Monomer
 Differences w/in IgG subclasses
are recognized by Gm allotypes.
 Subclasses also differ w/ respect to
binding of macrophages &
activation of complement.
 IgG

 Most serum IgG in NB is

maternal in origin.
 Capable of crossing placenta
 Accounts for 75% of total
serum Ig
 IgM

 Pentamer
 Itis the 1st Ab to appear after a
primary antigenic stimulus, the
1st to appear in phylogeny & the
last to leave in senescence.
 10% of total serum Ig
 IgM

 m.w. 900,000 dalton

 Most often formed in response to
stimulus by gram (-) bacteria.
 E.g. Wasserman Ab, heterophil Ab,
rheumatoid factor, cold agglutinins,
& allohemagglutinins
 IgM

 Concentration in cord serum is

fetal in origin & measures b/n 5
& 10% of that found in adult
serum.
 Has 2 subclasses: IgM1 & IgM2
(difference in μ chain)
 IgA

 Dimer
 Exists as serum IgA & secretory IgA
 Cannot be detected in cord serum
 Synthesized in plasma cells located
primarily in the epithelial surfaces of
the respiratory tract & intestine & in
almost all excretory glands.
 IgA

 activate
the alternative
pathway.
 IgE

 Originallycalled “reagin”
 Has role in allergic conditions
 cytotrophic
 IgD

 Monomer
 Heat & acid labile
 No proven antibody activity
 Predominant Ig on the surface
of B cells
IMMUNOGLOBULIN
STRUCTURE