Professional Documents
Culture Documents
and Function
Intended Learning Outcomes
● Differentiate and describe the five classes of immunoglobulins in
terms of structure, properties and functions.
● Correctly identify the parts and functions of the basic antibody
structure.
● Explain the different theories of antibody production
● Compare and contrast the primary and secondary responses to
antigen effectively.
● Accurately relate the influence of monoclonal antibodies to
current laboratory testing practices
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Overview:
● Antibodies are specific glycoproteins referred to as
immunoglobulins.
● They can be found in blood plasma, and in many body
fluids such as tears, saliva, and colostrum.
● The primary function of an antibody in the body
defenses is to combine with antigen, which can lead to
bacterial or viral neutralization.
● They are considered to be the main humoral element of
the adaptive immune response
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Immunoglobulins (Ig)
● Glycoprotein molecules which are produced by plasma cells in
response to an immunogen and which function as antibodies.
● The Immunoglobulins derive their name from the finding that when
antibody-containing serum is place in an electrical field, the
antibodies, which were responsible for immunity, migrated with
globular proteins.
● Immunoglobulins are divided into five major classes on the basis of a
part of the molecule called the heavy chain:
○ IgG, IgM, IgA, IgD, and IgE (with Ig being the abbreviation for
immunoglobulin)
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ANTIBODIES
1. Fixation of complement - This results in lysis of cells and release of
biologically active molecules
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Properties of an Antibody
1. Protein in nature
2. With high molecular weight
3. Present in serum/ plasma, saliva, semen,
CSF and other body fluids
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GENERAL FUNCTIONS OF
IMMUNOGLOBULINS
1. Neutralize toxic substances
2. Facilitate phagocytosis and kill microbes
3. Combine with antigens on cellular surfaces
and hereby cause the destruction of these
cells whether extravascularly or
intravascularly
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Classification of
Antibodies
Five classes of Antibodies:
IgG
IgA
IgM
IgD
IgE
Classification of antibodies
A. According to its sedimentation constant
SEDIMENTATION MOLECULAR WEIGHT
IMMUNGLOBULIN
COEFFICCIENT (in Daltons)
IgG 7s 150, 00
IgE 8s 190,000
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Classification of antibodies
B. According to temperature at which they react best:
1. Cold Antibodies
2. Warm Antibodies
C. According to occurrence
1. Natural antibodies
2. Immune antibodies
D. According to the species which produce them
1. Isoantibodies
2. Heterophile antibodies
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Classification of antibodies
E. According to its reaction with an antigen
1. Agglutinins 6. Opsonins
2. Precipitins 7. Neutralizing antibodies
3. Agglutinoids 8. Allergic antibodies
9. Antitoxins
4. Hemagglutinins
10. Complement fixing antibodies
5. Lysins
11. Blocking or Inhibitory
antibodies
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Classification of antibodies
F. According to their in vitro behavior
Complete Antibody Incomplete Antibody
Univalent; blocking;
Synonyms Bivalent; Saline acting
coagglutinating
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The Structure of Immunoglobulin
Monomer- basic structural unit of an antibody
Tetrapeptide made up of:
1. Heavy chain
○ With five principal antigenic types with their
corresponding Ig. (gamma, alpha, mu, delta, epsilon)
2. Light chain
○ With two antigenically defined types: kappa and
lambda
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Antibody Structure
A four-chain unit molecule, each L chain was bonded to an H
chain by means of an S–S bond and the H chains were joined
to each other by one or more S–S bonds
Disulfide bonds
These are chemical bonds essential for the normal 3 dimensional
structure of Ig
2 types of disulfide bonds:
1. Interchain
JMMC2. Intrachain
Antibody Structure
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JMMC Lifted from: Stevens, C. D. (2010). Clinical Immunology and Serology A Laboratory Perspective. Fourth Edition. F.A. Davis Company
Hinge region
○ Flexible part of the antibody located in the heavy chains.
○ Located between the CH1 and CH2 regions is known as the hinge region
○ It is more exposed to enzymes and chemical thus papain acts here to
produce Fab and Fc portions
○ The light chains are each linked to one-half of a heavy chain by
disulphide bonds at the proximal end
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Light Chains
● Analysis of several Bence Jones proteins revealed
that there were two main types of L chains, which
were designated as:
○ kappa (κ) chains and lambda (λ) chains
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Three-Dimensional
Structure of
Antibodies
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Three-Dimensional Structure
of Antibodies
● The basic four-chain structure of all
immunoglobulin molecules does not actually
exist as a straight Y shape.
● It is in fact, folded into compact globular
subunits based on the formation of balloon-
shaped loops at each of the domains
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Lifted from: Stevens, C. D. (2010). Clinical Immunology and Serology A Laboratory Perspective. Fourth Edition. F.A. Davis Company
DOMAINS
○ Are globular regions on polypeptide chain stabilized
by intrachain disulfide bonds
a. Domains on the heavy chain
● VH, CH1, CH2, CH3, (CH4)
b. Domains in the light chain
● VL, CL
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JMMC Lifted from: Stevens, C. D. (2010). Clinical Immunology and Serology A Laboratory Perspective. Fourth Edition. F.A. Davis Company
Regions on Polypeptide
Chain
Variable Region Constant Region
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Other Defintions
● Polymer
○ Ig composed of more than a single basic
monomeric unit
● J chain
○ Polypeptide chain which normally holds
polymeric Immunoglobulins
● Secretory component
○ A substance attached to polymeric IgA
found on secretions.
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Enzyme Digestion
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Enzyme Digestion
1. Cleavage with papain enzymes
○ Digestion with papain breaks the immunoglobulin molecule in
the hinge region before the H-H inter-chain disulfide bond.
○ This results in the formation of two identical fragments that
contain the light chain and the VH and CH1 domains of the heavy
chain.
○ Antigen binding - These fragments were called the Fab
fragments because they contained the antigen binding sites of
the antibody.
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Cleavage with papain
enzymes
JMMC Lifted from: Stevens, C. D. (2010). Clinical Immunology and Serology A Laboratory Perspective. Fourth Edition. F.A. Davis Company
Cleavage with papain enzymes
● Digestion with papain also produces a fragment that contains
the remainder of the two heavy chains each containing a CH2
and CH3 domain. This fragment was called Fc because it was
easily crystallized
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Enzyme Digestion
1. Cleavage with pepsin enzymes
○ Treatment of immunoglobulins with pepsin results in cleavage of
the heavy chain after the H-H inter-chain disulfide bonds resulting
in a fragment that contains both antigen binding sites
○ This fragment was called F(ab')2 because it is divalent.
○ The Fc region of the molecule is digested into small peptides by
pepsin.
○ The F(ab')2 binds antigen but it does not mediate the effector
functions of antibodies
3. Reduction using mercaptoethylamine
○ Break the disulfide bonds
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Cleavage with pepsin
enzymes
JMMC Lifted from: Stevens, C. D. (2010). Clinical Immunology and Serology A Laboratory Perspective. Fourth Edition. F.A. Davis Company
Enzyme
Digestion
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Characteristics of the Different
Types of Immunoglobulins
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Immunoglobulin
Classes
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IgG
● Structure: Monomer
● Percentage serum antibodies: 75- 75% of the total Immunoglobulin
pool
● Has 4 major subclasses: IgG1, IgG2, IgG3, IgG4
● Equally distributed in the different fluid compartments with detectable
amounts in CSF and urine
● IgG antibody response appears later than IgM in primary response but
they form the major antibody of the secondary immune response
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IgG
● Maternal IgG is actively and selectively transferred across the
placenta to the fetus and imparts passive protection to the newbon
for 6-9 months
● Functions of IgG:
○ Provides immunity for ,the newborn
○ Complement fixation
○ Opsonization
○ Neutralization of toxins and viruses
○ Participation in agglutination and precipitation reactions
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IgM
● Structure: Pentamer
● Percentage serum antibodies: 5-10% of the total
immunoglobulin pool
● Star-shaped in the free state; reaction with 10 functional
binding sites.
● crab-like in antigen-antibody Location: Blood and lymph
(pentamer), B cell surface (monomer)
● Half-life in serum: 6 days
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IgM
● A powerful agglutinator of a particulate antigen
● Maternal IgM does not cross the placenta
● Functions of IgM:
○ First antibodies produced during an infection but it does not
persist for long.
○ Complement fixation
○ Agglutination
○ Opsonization
○ Neutralization of toxins
○
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Surface receptor for antigens (on B cells)
IgM Response
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IgA
Structure: Dimer
● Percentage serum antibodies: 10-15% of human serum Ig pool
● Found in serum in small amounts but predominant in sero-mucous
secretions of the respiratory tract, genito-urinary tract and GI tracts
● Secretions: (tears, sweat, colostrum, saliva and breastmilk), blood and
lymph.
● Half-life in serum: 6 days
● Complement Fixation: No
●
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Placental Transfer: No
IgA
● Forms of IgA:
Serum IgA
- Can agglutinate motile infectious agents thus promoting their phagocytosis
but they cannot activate the complement system
Secretory IgA
- A polymeric form stabilized a short polypeptide chain.
- It is known as the “antiseptic paint” of mucous membranes.
- It can activate the bacteriolytic activtiy through the alternate pathway of
the complement system and only in the presence of lysozyme
● Known Functions: Localized protection of mucosal surfaces. Provides immunity to
infant digestive tract.
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IgD
● Structure: Monomer
● Percentage serum antibodies: 0.001% of total Ig pool, but
known to be present in large quantities on the membrane of
many circulating immunocompetent B lymphocytes and
unstimulated B cells
● Heat labile immunoglobulin
● Detected by highly sensitive assay requiring radio-labeled
antisera
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IgD
● Precise biological action is not known but it may play a role in
antigen-triggered lymphocyte differentiation
● Location: B-cell surface, blood, and lymph
● Half-life in serum: 3 days
● Complement Fixation: No
● Placental Transfer: No
● Known Functions: In serum function is unknown. On B cell
surface, initiate immune response
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IgE
● Structure: Monomer
● Percentage serum antibodies: 0.0005% of total serum Ig
● Most heat labile immunoglobulin
● Synthesized locally by plasma cell present in the mucous
membrane of the GI and respiratory tracts
● It is homocytropic due to its ability to attach to the human skin,
it is associated with immediate hypersensitivity reactions but
also, apparently, with immunity to certain helminthic parasites
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IgE
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IgE
● Also known as reaginic antibody/ nuissance antibody
● Location: Bound to mast cells and basophils throughout body.
Blood.
● Half-life in serum: 2 days
● Complement Fixation: No
● Placental Transfer: No
● Known Functions:
○ Allergic reactions. Possibly lysis of worms.
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Theories of Antibody
Production
Ehrlich’s Side Chain Theory
○ Certain cells has had specific surface receptors for antigen
that were present before contact with antigen occurred.
○ Once antigens was introduced, it would select the cell with
proper receptors, combination would take place and then
receptors will break off and enter the circulation as antibody
molecules.
○ New receptors will be formed in place of those broken off and
this process could be repeated.
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Theories of Antibody
Production
Clonal Selection Theory
○ Individual lymphocytes are gentically pre-programmed
to produce one type of immunoglobulin, and that
specific antigen finds or selects those particular cells
capable of responding to it, causing these to proliferate
○ Repeated contact with the antigen would continually
increase a lymphocyte pool
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jmcuevas@fatima.edu.ph
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JMMC 👍 Reminder: Be ready for an assessment task next meeting on Session 1.