Antigen-Antibody Interactions -
Antigen-antibody interactions depend on four types
of noncovalent interactions: hydrogen bonds, ionic
bonds, hydrophobic interactions, and van der Waals
interactions.
The affinity constant, which can be determined by
Scatchard analysis, provides a quantitative measure of the
strength of the interaction between an epitope of the antigen
and a single binding site of an antibody. The avidity reflects
the overall strength of the interactions between a
multivalent antibody molecule and a multivalent antigen
molecule at multiple sites.
The interaction of a soluble antigen and precipitating antibody
in a liquid or gel medium forms an Ag-Ab precipitate.
Electrophoresis can be combined with precipitation
in gels in a technique called immunoelectrophoresis.
The interaction between a particulate antigen and agglutinating
antibody (agglutinin) produces visible clumping, or
agglutination that forms the basis of simple, rapid, and
sensitive immunoassays.
Radioimmunoassay (RIA) is a highly sensitive and quantitative
procedure that utilizes radioactively labeled antigen
or antibody.
The enzyme-linked immunosorbent assay (ELISA) depends
on an enzyme-substrate reaction that generates a
colored reaction product. ELISA assays that employ
chemiluminescence instead of a chromogenic reaction are
the most sensitive immunoassays available.
In Western blotting, a protein mixture is separated by electrophoresis;
then the protein bands are electrophoretically
transferred onto nitrocellulose and identified with labeled
antibody or labeled antigen.
Fluorescence microscopy using antibodies labeled with
fluorescent molecules can be used to visualize antigen on
or within cells.
Flow cytometry provides an unusually powerful technology
for the quantitative analysis and sorting of cell populations
labeled with one or more fluorescent antibodies.
Antigen-Antibody Interactions -
Antigen-antibody interactions depend on four types
of noncovalent interactions: hydrogen bonds, ionic
bonds, hydrophobic interactions, and van der Waals
interactions.
The affinity constant, which can be determined by
Scatchard analysis, provides a quantitative measure of the
strength of the interaction between an epitope of the antigen
and a single binding site of an antibody. The avidity reflects
the overall strength of the interactions between a
multivalent antibody molecule and a multivalent antigen
molecule at multiple sites.
The interaction of a soluble antigen and precipitating antibody
in a liquid or gel medium forms an Ag-Ab precipitate.
Electrophoresis can be combined with precipitation
in gels in a technique called immunoelectrophoresis.
The interaction between a particulate antigen and agglutinating
antibody (agglutinin) produces visible clumping, or
agglutination that forms the basis of simple, rapid, and
sensitive immunoassays.
Radioimmunoassay (RIA) is a highly sensitive and quantitative
procedure that utilizes radioactively labeled antigen
or antibody.
The enzyme-linked immunosorbent assay (ELISA) depends
on an enzyme-substrate reaction that generates a
colored reaction product. ELISA assays that employ
chemiluminescence instead of a chromogenic reaction are
the most sensitive immunoassays available.
In Western blotting, a protein mixture is separated by electrophoresis;
then the protein bands are electrophoretically
transferred onto nitrocellulose and identified with labeled
antibody or labeled antigen.
Fluorescence microscopy using antibodies labeled with
fluorescent molecules can be used to visualize antigen on
or within cells.
Flow cytometry provides an unusually powerful technology
for the quantitative analysis and sorting of cell populations
labeled with one or more fluorescent antibodies.
Enzyme-Linked ImmunoSorbent Assay, or ELISA, is a biochemical technique used mainly in immunology to detect the presence of an antibody or an antigen in a sample. The ELISA has been used as a diagnostic tool in medicine and plant pathology, as well as a quality control check in various industries. In simple terms, in ELISA an unknown amount of antigen is affixed to a surface, and then a specific antibody is washed over the surface so that it can bind the antigen. This antibody is linked to an enzyme, and in the final step a substance is added that the enzyme can convert to some detectable signal.
Hypersenstivity type3 is an Immune-complex mediated hypersensitivity.Hypersensitivity denotes a condition in which an exaggerated immune response of a host to non-harmful antigens that leads to destruction of host tissues.
Antigen-Antibody Reaction (Ab Ag Reaction)PoojaVishnoi7
The ppt deals covers-
- A general introduction to what antigen antibody reaction is.
- Salient features of Antigen Antibody Reaction.
- Strength of Antigen Antibody Reaction.
- Types of Antigen-Antibody Reaction.
- Applications of Antigen Antibody Reactions
Immunity can be defined as a complex biological system endowed with the capacity to recognize and tolerate whatever belongs to the self, and to recognize and reject what is foreign.
Enzyme-Linked ImmunoSorbent Assay, or ELISA, is a biochemical technique used mainly in immunology to detect the presence of an antibody or an antigen in a sample. The ELISA has been used as a diagnostic tool in medicine and plant pathology, as well as a quality control check in various industries. In simple terms, in ELISA an unknown amount of antigen is affixed to a surface, and then a specific antibody is washed over the surface so that it can bind the antigen. This antibody is linked to an enzyme, and in the final step a substance is added that the enzyme can convert to some detectable signal.
Hypersenstivity type3 is an Immune-complex mediated hypersensitivity.Hypersensitivity denotes a condition in which an exaggerated immune response of a host to non-harmful antigens that leads to destruction of host tissues.
Antigen-Antibody Reaction (Ab Ag Reaction)PoojaVishnoi7
The ppt deals covers-
- A general introduction to what antigen antibody reaction is.
- Salient features of Antigen Antibody Reaction.
- Strength of Antigen Antibody Reaction.
- Types of Antigen-Antibody Reaction.
- Applications of Antigen Antibody Reactions
Immunity can be defined as a complex biological system endowed with the capacity to recognize and tolerate whatever belongs to the self, and to recognize and reject what is foreign.
Antigen ,Antibody and Ag-Ab reactions ppt by DR.C.P.PRINCEDR.PRINCE C P
An immunogen refers to a molecule that is capable of eliciting an immune response, whereas an antigen refers to a molecule that is capable of binding to the product of that immune response (Ab).
So, an immunogen is necessarily an antigen, but an antigen may not necessarily be an immunogen
The terms immunogen and antigen are often used interchangeably but the later is more common.
Antibodies are Globulin Protein (Immunoglobulin) that are synthesized in the Serum and Tissue fluids.
It reacts specifically with the antigen that stimulated their production.
There are two types serum proteins: albumin and globulin
There are Three types of globulins .
1. Alpha globulin
2. Beta globulin
3. Gamma globulin (Antibodies)
Gamma globulins are responsible for immunity. So they are called as Immunoglobulin (Ig)
The binding of an antibody with an antigen of the type that stimulated the formation of antibody that results in the following reaction
Agglutination
Precipitation
Complement fixation
Phagocytosis
Neutralization of an exotoxin
Opsonization
Tissue fixation
Chemotaxis
Activation of mast cells and basophils
PPT prepared by:
DR.PRINCE C P
Associate Professor , Department of Microbiology,
Mother Theresa Post Graduate & Research Institute of Health Sciences (Government of Puducherry Institution)
This topic covers the brief introduction of Ag and Ab in detail. Types and functions of Ig is explained in detail. Paraproteinemias is explained with simple pictures.
by Dr. N.Sivaranjani, MD
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Immune Response: Third line of defense. Involves production of antibodies and generation of specialized lymphocytes against specific antigens.
Antigen: Molecules from a pathogen or foreign organism that provoke a specific immune response
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2. Antigen
Any substance which ,when introduced
parentrally into the body, stimulates
the production of antibody with which
it reacts specifically & in observable
manner.
Either humoral or cell mediated
immunity
3.
4. Specificity – Ag introduced in body
react with those particular
immunocytes which carry specific
marker for that antigen
Ab produced will react with that
particular Ag
5. Epitope
Smallest unit of antigenicity, antigenic
determinant
Small area on Ag capable of combining
complementary site on Ab or T
lymphocyte.
9. 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.
Lipids and nucleic acids are only antigenic
when combined with proteins or
polysaccharides.
10. Antibody
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.
Valency of an antibody: Number of antigen
binding sites. Most are bivalent.
Belong to a group of serum proteins called
immunoglobulins (Igs).
11. Function of Ab
Elimination of foreign antigen
Recruitment& enhancement of host
effector mechanism
antibody levels in diagnosis
Passive administration -therapy
15. Antibody structure
Monomer: A flexible Y-shaped molecule with
four protein chains:
– 2 identical light chains
– 2 identical heavy chains
Variable Regions: Two sections at the end of Y’s
arms. Contain the antigen binding sites (Fab).
Identical on the same antibody, but vary from
one antibody to another.
Constant Regions: Stem of monomer and lower
parts of Y arms.
Fc region: Stem of monomer only. Important
because they can bind to complement or cells.
16. Lock and Key Concept
concept of antigen-antibody reactions
is one of a key (i.e. the antigen) which
fits into a lock (i.e. the antibody).
Ab
Ag
18. Immunoglobulin –proteins of
animal origin
Chemically antibodies are
globulines and hence they are
called as immunoglobulines
Synthesised by plasma cells & B
lymphocytes
All antibodies are Ig. But not true
vice-versa
21. IgG
Structure: Monomer
Percentage serum antibodies:
80%Most common
Location: Blood(12mg/ml), lymph, intestine
Molecular weight150 kDa
Half-life in serum: 23 days
22. IgG
Complement Fixation, Neutralisation,
precipitation : Yes
Diffuses easily IV & EV compartments
Placental Transfer: Yes
Known Functions: Enhances
phagocytosis, neutralizes toxins and
viruses, protects fetus and newborn.
23. IgG
Catabolic rate depends directly on
conc. Of IgG.
Passive immunisation suppresses
homologus Ab producion used in
Anti RhiD isoimmunisation
IgG1, IgG2,IgG3,IgG4
IgG1, IgG3 activators of classical
complement pathway
25. IgM
Structure: Pentamer
Percentage serum antibodies: 5-10%
Location: Blood(0.5-2mg/100ml), lymph, B
cell surface (monomer)
Half-life in serum: 5 days
Complement Fixation: Yes
Placental Transfer: No
Known Functions: First antibodies
produced during an infection. Effective
against microbes and agglutinating
antigens.
26. Pentamer, 5 H & L chains, One
molecule of J Chain
Can not spread from blood to
tissues
27. Functions of IgM
Protection against bacteraemia
Pramotes phagocytosis &
bacteriolysis by complement
activation
Primary immune response
infection
Can not cross placenta- presence
in new borne IU infection
Major component of rheumatoid
factor
30. IgA
Structure: Dimer
Percentage serum antibodies: 10-15%
Location: Secretions (tears, saliva,
intestine, milk), blood and lymph.
Half-life in serum: 6 days
Complement Fixation: No
Placental Transfer: No
Known Functions: Localized protection of
mucosal surfaces. Provides immunity to
infant digestive tract.
31. Serum IgA & Secretory IgA
Covers microorganisms and
prevents adhesion to mucosal
surface
33. IgD
Structure: Monomer
Percentage serum antibodies: 0.2%
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.
35. IgE
Structure: Monomer
Percentage serum antibodies:
0.002%
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.
36. It is mostly distributed in
extravascular space.
IgE + mast cell serves as receptor
for allergens and parasitic antigens
Mast cells degranulated with
release of vasoactive amines
Increased IgE parasitic infection
38. Multiple myeloma
Plasma cell tumor in bone marrow
Producing excess of single class
of immunoglobulin (M proteins)
All 5 classes
Myeloma with IgM producing cells
is called as Waldenstrom’s
macroglobulinemia
39. Bence Jones protein
Light chains of immunoglobulins
Present in urine of myeloma
patients
Either kappa or lambda
Coagulates when heated at 600
C
redisolves at 800
C
40. Cryoglobulinaemia
Presence of cryoglobulin in blood
Precipitate in microvasculature on
exposure to cold
Associated with SLE, myeloma,
macroglobulinaemia.
Made up of IgG, IgM or mixture