LECTURE NOTESANTIBODY STRUCTURE AND FUNCTION

immunoglobulins (Ig): glycoproteins found in the serum portion of the blood

when subjected to electrophoresis at pH 8.6, Ig appear primarily in gamma () band

Ig classesIgA, IgD, IgE, IgG & IgM

IMMUNOGLOBULIN STRUCTURE

basic unit of all Ig classes is composed of 4 polypeptide chains

2 heavy (H) chains
2 light (L) chains

basic structure of Ig was resolved by experiments using proteolytic enzymes

papaincleaves Ig into 3 fragments (2 Fab fragments and 1 Fc fragment)
pepsincleaves Ig into 2 fragments (1 F(ab)2 fragment and 1 Fc fragment)
Light Chains
Bence-Jones proteinsfound in urine of patients with multiple myeloma; represent L chains
that were being secreted by the malignant plasma cells
analysis revealed there were two types of L chains: kappa () and lambda ()

Heavy Chains
constant region of H chains unique for each class of Ig and gives each Ig its name (i.e.
isotype)

allotype: minor sequence variations in the constant region of H or L chains within a given
class of Ig or L chain
IgG allotypes
IgA allotypes
-L chain allotypes

idiotype: variable portions of each chain are unique to a specific antibody molecule; present
at the amino-terminal end of the H and L chains
forms the antigen binding region of Ig

hinge region: proximal portion of the H chain constant region; has high content of proline
and hydrophobic amino acid residues
proline residues provide flexibility to Ig tertiary structure
permits Ag-binding sites to operate independently
enhances effector functions of Ig (e.g. initiation of complement cascade)

TABLE 4-1 (Stevens, p. 59)

Table 4-1 Properties of Immunoglobulins

IgG IgM IgA IgD IgE

Molecular weight 150,000 900,000 160,000- 180,000 190,000

H chain

H chain subclasses 1, 2, 3, 4 None 1, 2 None None

Serum concentration (mg/dl) 800-1600 120-150 70-350 1-3 0.005

Complement fixation Yes Yes No No No

Crosses placenta Yes No No No No

IgG
 provides immunity for newborns
classical pathway complement activation
opsonization (neutrophils, monocytes, macrophages have Fc-receptors for IgG)
neutralizing toxins
neutralizing viruses
participation in agglutination and precipitation reactions

IgM
due to large size, found mainly in the intravascular pool (serum) not in other body fluids or
tissue
first antibody to appear following antigen stimulation (early antibody)
no memory B-cells generated that produce IgM (due Ig class switching)
most efficient activator of classical pathway complement activation
participates in agglutination reactions
opsonization
toxin neutralization

IgA
major Ig in secretions
exist as a dimer along respiratory, urogenital, and intestinal mucosa
also present in milk, saliva, tears and sweat
secretory component (SC) attached to Fc portions of Ig unit
SC facilitates transport of IgA to mucosal surfaces
SC makes IgA resistant to proteolytic digestion
first line of specific defense
neutralizes toxins produced by microorganisms
prevents bacterial adherence to mucosal surfaces
IgA immune complexes activate alternative pathway complement activation
Fc-receptors for IgA present on neutrophils, monocyte, macrophagesbinding of receptor
triggers respiratory burst and degranulation during phagocytosis

IgD
primarily present on the surface of immunocompetent nave (unstimulated) B-cells
second Ig produced by nave B-cells (IgM produced first)
plays a role in B-cell activation
may play a role in B-cell maturation and differentiation

IgE
least abundant Ig in serum
shortly after synthesis, attaches to basophils and tissue mast cells
high-affinity Fc-receptors for IgE present on basophils and mast cells
 binding and cross-linking of antigen triggers basophil/mast cell degranulation and release
of vasoactive amines (histamine and heparin)type I hypersensitivity reaction (i.e. allergic
response)
allergic response may be important in attracting neutrophils and eosinophils during acute
inflammation
eosinophils important for the destruction of large antigens such as parasitic worms that
cannot be easily phagocytosed

MONOCLONAL ANTIBODY PRODUCTION

A technique to produce antibody from a single B-cell.

Developed by Kohler and Milstein in 1975 (Nobel Prize 1984).

Fused activated B-cells with myeloma cells incapable of producing Ig.

CLINICAL APPLICATIONS OF MONOCLONAL ANTIBODIES

monoclonal antibodies used for in vitro diagnostic testing (numerous laboratory reagents)

monoclonal antibodies use as therapeutic agents

autoimmune diseasesrheumatoid arthritis, Crohns disease (e.g. Humira &
Ratuxan)
cancermetastatic breast cancer, lymphomas, colorectal cancer, head & neck cancers,
non-small cell lung cancer