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Antibody (Reuploaded)
Antibody (Reuploaded)
Antibody (Reuploaded)
Antibodies (Immunoglobulins)
bifunctional glycoproteins serum portion of the blood gamma band
Structure of Immunoglobulins
basic structural units - monomer four chain of polypeptide unit
Light or L chains (200 amino acids & 25,000 daltons) Heavy or H chains (2x the light chain)
noncovalent forces disulphide interchain bridges Hinge region location between CH1 and CH2 high proline content - flexibility hydrophobic residues
Carbohydrate portions
localized between CH2 domains
functions:
increase solubility of the Igs provide protection against degradation
Variable region
Differ between Igs Antigen binding fragment (Fab) Determines specificity Amino terminal end (NH2) Amino acid sequence is changing Binds to antigens
Constant region
Shared by many Igs Crystallizable fragment (Fc)
Binds to effector cells Carboxy terminal end (COOH) Amino acid sequence is fixed
Light Chains
kappa& lambda chains carboxy terminal ends differ amino acid substitution both present in all classes but only
Heavy chains
5 classes of Igs w/ different constant regions alpha, delta, gamma, mu, epsilon
Domains
globular regions disulphide intrachain bridges light chains VL & CL heavy chains VL & CH1, CH2 & CH3
Domain
VL & VH CH1 Antigen binding
Functions
CH2
CH3
Papain cleavage
amino terminal side of the disulphide bridges 2 identical fragments w/ antigen binding activity
Pepsin cleavage
carboxy terminal side of the disulphide bridges F(ab)2 (2 arms remain linked)
several Fc portions
Fd
papain digestion & reduction w/ mercaptoethylamine amino terminal half (heavy chain)
Properties of Ig Isotypes IgG blood & interstitial fluid transported across the placenta major role in elimination of micobes opsonization C activation ADCC neutralization
IgM
Macroglobulin mol. wt. of 990,000 5 monomeric units held by a J chain Assumes a star-like shape 3 dimensional structure crab-like mainly in intravascular pool can not cross the placenta first to appear in the primary response no memory cell for IgM IgM receptor mature B cells
functions
most efficient in triggering the classical C pathway powerful agglutinator involves in opsonization useful indicator of intrauterine infection
IgA
10 to 15% of the Ig pool
IgD
less than 0.2% in the serum extended hinge region
disulphide bonds
IgE
0.004% not capable of crossing the placenta heat-labile --- conformational changes no binding to target cells does not participate in typical Ig reactions
Feature shared by IgA, IgM and IgD carboxy terminal octa-deca peptide tail CH4 IgM and IgE
Allotype
unique differences
within the constant regions gamma & alpha heavy chains kappa light chains present in different individuals IgG Gm designation IgA Am designation Kappa light chain Km designation
Idiotype
Clonal Selection Theory Lymphocytes are generally preendowed to respond to one antigen or group of antigens. That IgM &IgD act as surface receptors that interact w/ specific antigen to trigger proliferation of a clone of identical cells.
(gene coding for Igs) 1. Separate diversity exists for each chain since they are coded for on separate chromosomes Heavy Chain Chromosome 14
Gene organization (gene coding for Igs) 2. Analysis of Ig genes has revealed that the variable & constant regions are separately encoded & located on different fragments of DNA 3. Four separate chromosomes code for heavy chains, three separate chromosomes code for light chains Heavy Chain VDJC Light Chain - VJC
a. During the process of B cell maturation, the pieces are sliced together to commit the B cell to make Ab of single specificity b. Joining of the segments occurs in two steps: 1) at the DNA level, 1D gene is joined to the J gene, with deletion of intervening DNA 2) the V gene is joined to the DJ complex resulting in rearranged VDJ complex. This results in pro B cell.
c. When RNA synthesis occurs, C gene is joined to the VDJ complex & all intervening DNA is lost. In general, the C gene which is immediately located next to the VDJ complex is expressed. Since C mu gene is the closest to the J segment, it is the Ab that is recognized d. W/ the exception of C delta gene which lies next to the C mu gene is often transcribed along with the C mu gene. Thus, B cell will have IgM & IgD at the same variable domain on the surface membrane at the same time The process of switching to other classes of Ig occurs later due to the looping out & deletion of other constant regions
A productive rearrangement of the kappa chains keeps other chromosome 2 from rearranging. In addition to this, it shuts off any recombination of lambda chain locus on chromosome 22. This process is known as allelic exclusion.
Lambda chain synthesis occurs only if nonfunctional gene product arises from kappa chains The light chain is joined to the mu chain to form a complete IgM Ab which appears first in immature B cell. Once the IgD & IgM are present on the surface membrane of B cell, the B cell is fully mature & capable of responding to an antigen.
The large variety of VDJ & C combinations for each type of chain plus the different possibilities for light & heavy chain make more than enough configurations that allow us to respond to any antigen in the environment. Genetic preprogramming of lymphocytes can best be explained by the concept of gene recombination. More than 1 gene controls synthesis of a particular Ig and through a random selection process these individual segments are joined to commit that lymphocyte to making Ab of a single specificity.
HAT medium
Classification 1. Sedimentation constant IgM IgA IgE Serum IgA 19S 13S, 11S, 9S 8S 7S
IgG
IgD
7S
7S
2. Temperature
Cold 4oC or RT Warm 37oC
3. Occurrence
Natural Immune
lysin
opsonin blocking or inhibitory antibodies complement fixing antibodies
6. In vitro behaviour Complete thermolabile cant cross the placenta early in occurrence saline acting Incomplete thermostable can cross the placenta late in occurrence albumin acting
The half life of most IgG subclasses is approximately 23 days. True or False
Some of the antibodies in the serum from a blood group type A individual will bind to RBCs from an individual who is type O. True or False
Treatment of antibody molecule w/ papain generates two Fab fragments and one Fc fragment. True or False
True or False
Plasma cells derived from one B cell clone, secrete antibodies that all recognize the same epitope. True or False
The antibody isotype (when complexed with antigen) that serves as an opsonin for phagocytes is of the __________isotype. IgG
IgM