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IMMUNOLOGY - CHAPTER FOUR

IMMUNOGLOBULINS - STRUCTURE AND FUNCTION
Dr. Gene Mayer

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Logo image Jeffrey Nelson, Rush

University, Chicago, Illinois and The
MicrobeLibrary

READING
Male et al. Immunology
7th edition Chapter 3
Murray, et al. Medical Microbiology
5th edition, pp 110-113
TEACHING OBJECTIVES

I. DEFINITION
Immunoglobulin (Ig)
Immunoglobulins are glycoprotein molecules that are produced by plasma cells in response to an immunogen and which function as
antibodies. The immunoglobulins derive their name from the finding that they migrate with globular proteins when antibody-containing serum is
placed in an electrical field (Figure 1).

To discuss the general properties of all

immunoglobulins
To describe the basic structure of immunoglobulins
To relate immunoglobulin structure with function
To define immunoglobulin hypervariable and
framework regions

II. GENERAL FUNCTIONS OF IMMUNOGLOBULINS

A. Antigen binding
Immunoglobulins bind specifically to one or a few closely related antigens. Each immunoglobulin actually binds to a specific antigenic
determinant. Antigen binding by antibodies is the primary function of antibodies and can result in protection of the host. The valency of
antibody refers to the number of antigenic determinants that an individual antibody molecule can bind. The valency of all antibodies is at least
two and in some instances more.

B. Effector Functions
To define immunoglobulin classes and subclasses, Frequently the binding of an antibody to an antigen has no direct biological effect. Rather, the significant biological effects are a consequence
types and subtypes
of secondary "effector functions" of antibodies. The immunoglobulins mediate a variety of these effector functions. Usually the ability to carry
out a particular effector function requires that the antibody bind to its antigen. Not every immunoglobulin will mediate all effector functions.
To describe the structures and properties of
Such effector functions include:
immunoglobulin classes

1. Fixation of complement - This results in lysis of cells and release of biologically active molecules (see chapter two)

Figure 1 Electrophoretic
separation of serum proteins

KEY WORDS
Immunoglobulin
Valence
Heavy chain

2. Binding to various cell types - Phagocytic cells, lymphocytes, platelets, mast cells, and basophils have receptors that bind immunoglobulins.
This binding can activate the cells to perform some function. Some immunoglobulins also bind to receptors on placental trophoblasts, which
results in transfer of the immunoglobulin across the placenta. As a result, the transferred maternal antibodies provide immunity to the fetus and
newborn

III. BASIC STRUCTURE OF IMMUNOGLOBULINS

The basic structure of the immunoglobulins is illustrated in figure 2. Although different immunoglobulins can differ structurally, they all are built

Light chain
Variable region
Constant region
Hinge region
Domain
Hypervariable region
Framework region
Groups & subgroups
Fab & Fc, F(ab')2
Type & subtype
Class & subclass
Opsonin
J chain
Secretory component

from the same basic units.

A. Heavy and Light Chains
All immunoglobulins have a four chain structure as their basic unit. They are composed of two identical light chains (23kD) and two identical
heavy chains (50-70kD)
B. Disulfide bonds
1. Inter-chain disulfide bonds - The heavy and light chains and the two heavy chains are held together by inter-chain disulfide bonds and by
non-covalent interactions The number of inter-chain disulfide bonds varies among different immunoglobulin molecules.
2. Intra-chain disulfide binds - Within each of the polypeptide chains there are also intra-chain disulfide bonds.
C. Variable (V) and Constant (C) Regions
When the amino acid sequences of many different heavy chains and light chains were compared, it became clear that both the heavy and light
chain could be divided into two regions based on variability in the amino acid sequences. These are the:
1. Light Chain - VL (110 amino acids) and CL (110 amino acids)
2. Heavy Chain - VH (110 amino acids) and CH (330-440 amino acids)

Figure 2A The basic

structure of immunoglobulins

Figure 2B
Click on the image at left for an animated tutorial
on antibody structure Requires Chime Plug-In. Get Chime
here. Developed by Eric Martz. Development supported by the
Division of Undergraduate Education of the National Science

D. Hinge Region
This is the region at which the arms of the antibody molecule forms a Y. It is called the hinge region because there is some flexibility in the
molecule at this point.
E. Domains
Three dimensional images of the immunoglobulin molecule show that it is not straight as depicted in figure 2A. Rather, it is folded into globular
regions each of which contains an intra-chain disulfide bond (figure 2B-D). These regions are called domains.
1. Light Chain Domains - VL and CL

2. Heavy Chain Domains - VH, CH1 - CH3 (or CH4)

F. Oligosaccharides
Carbohydrates are attached to the CH2 domain in most immunoglobulins. However, in some cases carbohydrates may also be attached at other
locations.

Foundation.

Figure 2C Ribbon drawing of

the first intact antibody (IgG2A) every crystallized.
Harris, L. J., Larson, S. B., Hasel, K. W., Day, J., Greenwood, A.,
McPherson, A. Nature 1992, 360, 369-372. 2000 Antibody
Resource Page

IV. STRUCTURE OF THE VARIABLE REGION

A. Hypervariable (HVR) or complementarity determining regions (CDR)
Comparisons of the amino acid sequences of the variable regions of immunoglobulins show that most of the variability resides in three regions
called the hypervariable regions or the complementarity determining regions as illustrated in figure 3. Antibodies with different specificities (i.e.
different combining sites) have different complementarity determining regions while antibodies of the exact same specificity have identical
complementarity determining regions (i.e. CDR is the antibody combining site). Complementarity determining regions are found in both the H
and the L chains.
B. Framework regions

Figure 2D
Rotating antibody

Jose Saldanha, Humanization by Design

2000, Antibody Resource Page

The regions between the complementarity determining regions in the variable region are called the framework regions (figure 3). Based on
similarities and differences in the framework regions the immunoglobulin heavy and light chain variable regions can be divided into groups and
subgroups. These represent the products of different variable region genes.

V. IMMUNOGLOBULIN FRAGMENTS: STRUCTURE/FUNCTION RELATIONSHIPS

Immunoglobulin fragments produced by proteolytic digestion have proven very useful in elucidating structure/function relationships in
Figure 3 Structure of the

variable region framework regions

Click on image at left to see a

rotable molecular structure of an Fab fragment
bound to an influenza hemagglutinin peptide.
Requires Chime plug-in. Get Chime here)

Click on image at left to see

details of the interaction of a mouse monoclonal
antibody interacting with hen egg white lysozyme.

immunoglobulins.
A. Fab
Digestion with papain breaks the immunoglobulin molecule in the hinge region before the H-H inter-chain disulfide bond Figure 4. This results
in the formation of two identical fragments that contain the light chain and the V H 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. Each Fab
fragment is monovalent whereas the original molecule was divalent. The combining site of the antibody is created by both V H and VL. An
antibody is able to bind a particular antigenic determinant because it has a particular combination of V H and VL. Different combinations of a VH
and VL result in antibodies that can bind a different antigenic determinants.
B. Fc
Digestion with papain also produces a fragment that contains the remainder of the two heavy chains each containing a C H2 and CH3 domain.
This fragment was called Fc because it was easily crystallized.

Requires Chime plug-in. Get Chime here)

Figure 4 Immunoglobulin
fragments: Structure/function relationships

Figure 5 Immunoglobulin
fragments: Structure function relationships

Effector functions - The effector functions of immunoglobulins are mediated by this part of the molecule. Different functions are mediated by the
different domains in this fragment (figure 5). Normally the ability of an antibody to carry out an effector function requires the prior binding of an
antigen; however, there are exceptions to this rule.

C. F(ab')2
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 (figure 6). 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.

VI. HUMAN IMMUNOGLOBULIN CLASSES, SUBCLASSES, TYPES AND SUBTYPES

Figure 6 Immunoglobulin
fragments: Structure/function relationships

A. Immunoglobulin classes
The immunoglobulins can be divided into five different classes, based on differences in the amino acid sequences in the constant region of the
heavy chains. All immunoglobulins within a given class will have very similar heavy chain constant regions. These differences can be detected
by sequence studies or more commonly by serological means (i.e. by the use of antibodies directed to these differences).
1. IgG - Gamma heavy chains
2. IgM - Mu heavy chains
3. IgA - Alpha heavy chains
4. IgD - Delta heavy chains
5. IgE - Epsilon heavy chains
B. Immunoglobulin Subclasses
The classes of immunoglobulins can de divided into subclasses based on small differences in the amino acid sequences in the constant region
of the heavy chains. All immunoglobulins within a subclass will have very similar heavy chain constant region amino acid sequences. Again
these differences are most commonly detected by serological means.
1. IgG Subclasses
a) IgG1 - Gamma 1 heavy chains

b) IgG2 - Gamma 2 heavy chains

c) IgG3 - Gamma 3 heavy chains
d) IgG4 - Gamma 4 heavy chains
2. IgA Subclasses
a) IgA1 - Alpha 1 heavy chains
b) IgA2 - Alpha 2 heavy chains

C. Immunoglobulin Types
Immunoglobulins can also be classified by the type of light chain that they have. Light chain types are based on differences in the amino acid
sequence in the constant region of the light chain. These differences are detected by serological means.
1. Kappa light chains
2. Lambda light chains
D. Immunoglobulin Subtypes
The light chains can also be divided into subtypes based on differences in the amino acid sequences in the constant region of the light chain.
1. Lambda subtypes
a) Lambda 1

b) Lambda 2
c) Lambda 3
d) Lambda 4
E. Nomenclature
Immunoglobulins are named based on the class, or subclass of the heavy chain and type or subtype of light chain. Unless it is stated precisely,
you should assume that all subclass, types and subtypes are present. IgG means that all subclasses and types are present.
F. Heterogeneity
Immunoglobulins considered as a population of molecules are normally very heterogeneous because they are composed of different classes
and subclasses each of which has different types and subtypes of light chains. In addition, different immunoglobulin molecules can have
different antigen binding properties because of different V H and VL regions.

VII. STRUCTURE AND SOME PROPERTIES OF IG CLASSES AND SUBCLASSES

Figure 7 IgG Structure

A. IgG
1. Structure
The structures of the IgG subclasses are presented in figure 7. All IgG's are monomers (7S immunoglobulin). The subclasses differ in the
number of disulfide bonds and length of the hinge region.
2. Properties
IgG is the most versatile immunoglobulin because it is capable of carrying out all of the functions of immunoglobulin molecules.

a) IgG is the major Ig in serum - 75% of serum Ig is IgG

b) IgG is the major Ig in extra vascular spaces
c) Placental transfer - IgG is the only class of Ig that crosses the placenta. Transfer is mediated by a receptor on placental cells for the Fc
region of IgG. Not all subclasses cross equally well; IgG2 does not cross well.
d) Fixes complement - Not all subclasses fix equally well; IgG4 does not fix complement
e) Binding to cells - Macrophages, monocytes, PMNs and some lymphocytes have Fc receptors for the Fc region of IgG. Not all subclasses
bind equally well; IgG2 and IgG4 do not bind to Fc receptors. A consequence of binding to the Fc receptors on PMNs, monocytes and
macrophages is that the cell can now internalize the antigen better. The antibody has prepared the antigen for eating by the phagocytic cells.
The term opsonin is used to describe substances that enhance phagocytosis. IgG is a good opsonin. Binding of IgG to Fc receptors on other
types of cells results in the activation of other functions.

B. IgM
Figure 8 Pentameric
serum IgM structure

1. Structure
The structure of IgM is presented in figure 8. IgM normally exists as a pentamer (19S immunoglobulin) but it can also exist as a monomer. In
the pentameric form all heavy chains are identical and all light chains are identical. Thus, the valence is theoretically 10. IgM has an extra
domain on the mu chain (CH4) and it has another protein covalently bound via a S-S bond called the J chain. This chain functions in
polymerization of the molecule into a pentamer.

Figure 9 Cell surface IgM 2. Properties

structure

a) IgM is the third most common serum Ig.

b) IgM is the first Ig to be made by the fetus and the first Ig to be made by a virgin B cells when it is stimulated by antigen.
c) As a consequence of its pentameric structure, IgM is a good complement fixing Ig. Thus, IgM antibodies are very efficient in leading to the
lysis of microorganisms.
d) As a consequence of its structure, IgM is also a good agglutinating Ig . Thus, IgM antibodies are very good in clumping microorganisms for
eventual elimination from the body.
e) IgM binds to some cells via Fc receptors.
Figure 10 B cell antigen
receptor (BcR)

f) B cell surface Ig
Surface IgM exists as a monomer and lacks J chain but it has an extra 20 amino acids at the C-terminus to anchor it into the membrane (figure
9). Cell surface IgM functions as a receptor for antigen on B cells. Surface IgM is noncovalently associated with two additional proteins in the
membrane of the B cell called Ig-alpha and Ig-beta as indicated in figure 10. These additional proteins act as signal transducing molecules
since the cytoplasmic tail of the Ig molecule itself is too short to transduce a signal. Contact between surface immunoglobulin and an antigen is
required before a signal can be transduced by the Ig-alpha and Ig-beta chains. In the case of T-independent antigens, contact between the
antigen and surface immunoglobulin is sufficient to activate B cells to differentiate into antibody secreting plasma cells. However, for Tdependent antigens, a second signal provided by helper T cells is required before B cells are activated.

C. IgA

Figure 11 IgA Structure

1. Structure
Serum IgA is a monomer but IgA found in secretions is a dimer as presented in Figure 11. When IgA exits as a dimer, a J chain is associated
with it.
When IgA is found in secretions is also has another protein associated with it called the secretory piece or T piece; sIgA is sometimes referred
to as 11S immunoglobulin. Unlike the remainder of the IgA which is made in the plasma cell, the secretory piece is made in epithelial cells and

is added to the IgA as it passes into the secretions (Figure 12). The secretory piece helps IgA to be transported across mucosa and also
protects it from degradation in the secretions.
2. Properties
a) IgA is the 2nd most common serum Ig.

Figure 12 Origin of soluble IgA

b) IgA is the major class of Ig in secretions - tears, saliva, colostrum, mucus. Since it is found in secretions secretory IgA is important in local
(mucosal) immunity.
c) Normally IgA does not fix complement, unless aggregated.
d) IgA can binding to some cells - PMN's and some lymphocytes.

D. IgD
Figure 13 IgD Structure
1. Structure
The structure of IgD is presented in the Figure 13. IgD exists only as a monomer.
2. Properties
a) IgD is found in low levels in serum; its role in serum uncertain.
b) IgD is primarily found on B cell surfaces where it functions as a receptor for antigen. IgD on the surface of B cells has extra amino acids at
C-terminal end for anchoring to the membrane. It also associates with the Ig-alpha and Ig-beta chains.
c) IgD does not bind complement.

E. IgE
1. Structure
The structure of IgE is presented in Figure 14. IgE exists as a monomer and has an extra domain in the constant region.
2. Properties
a) IgE is the least common serum Ig since it binds very tightly to Fc receptors on basophils and mast cells even before interacting with antigen.
b) Involved in allergic reactions - As a consequence of its binding to basophils an mast cells, IgE is involved in allergic reactions. Binding of the
allergen to the IgE on the cells results in the release of various pharmacological mediators that result in allergic symptoms.
Figure 14

IgE Structure

c) IgE also plays a role in parasitic helminth diseases. Since serum IgE levels rise in parasitic diseases, measuring IgE levels is helpful in
diagnosing parasitic infections. Eosinophils have Fc receptors for IgE and binding of eosinophils to IgE-coated helminths results in killing of the
parasite.
d) IgE does not fix complement.

CLINICAL IMPLICATIONS OF HUMAN IMMUNOGLOBULIN CLASSES

Adapted from:F.T. Fischbach in "A Manual of Laboratory Diagnostic Tests," 2nd Ed., J.B. Lippincott Co., Philadelphia, PA, 1984.
Figure 15
Rotating antibody
Antibody Concepts

IgG

2000 Antibody Resource Page

1. Increases in:
a) Chronic granulomatous infections
b) Infections of all types
c) Hyperimmunization
d) Liver disease
e) Malnutrition (severe)
f) Dysproteinemia
g) Disease associated with hypersensitivity granulomas, dermatologic disorders, and IgG myeloma
h) Rheumatoid arthritis
2. Decreases in:
a) Agammaglobulinemia
b) Lymphoid aplasia
c) Selective IgG, IgA deficiency
d) IgA myeloma
e) Bence Jones proteinemia
f) Chronic lymphoblastic leukemia

IgM

1. Increases (in adults) in:

a) Waldenstrm's macroglobulinemia
b) Trypanosomiasis
c) Actinomycosis
d) Carrin's disease (bartonellosis)
e) Malaria
f) Infectious mononucleosis
g) Lupus erythematosus
h) Rheumatoid arthritis
I) Dysgammaglobulinemia (certain cases)
Note: In the newborn, a level of IgM above 20 ng./dl is an indication of in utero stimulation of the immune system and stimulation by the rubella
virus, the cytomegalovirus, syphilis, or toxoplasmosis.
2. Decreases in:
a) Agammaglobulinemia
b) Lymphoproliferative disorders (certain cases)
c) Lymphoid aplasia
d) IgG and IgA myeloma
e) Dysgammaglobulinemia
f) Chronic lymphoblastic leukemia

IgA
1. Increases in:
a) Wiskott-Aldrich syndrome

b) Cirrhosis of the liver (most cases)

c) Certain stages of collagen and other autoimmune disorders such as rheumatoid arthritis and lupus erythematosus
d) Chronic infections not based on immunologic deficiencies
e) IgA myeloma
2. Decreases in:
a) Hereditary ataxia telangiectasia
b) Immunologic deficiency states (e.g., dysgammaglobulinemia, congenital and acquired agammaglobulinemia, and hypogammaglobulinemia)
c) Malabsorption syndromes
d) Lymphoid aplasia
e) IgG myeloma
f) Acute lymphoblastic leukemia
g) Chronic lymphoblastic leukemia

IgD
1. Increases in:
a) Chronic infections
b) IgD myelomas

IgE
1. Increases in:
a) Atopic skin diseases such as eczema
b) Hay fever
c) Asthma
d) Anaphylactic shock

e) IgE-myeloma
2. Decreases in:
a) Congenital agammaglobulinemia
b) Hypogammaglobulinemia due to faulty metabolism or synthesis of immunoglobulins

Antibody Structure and ClassificationNote 7.1

The basic structural unit of most mammalian antibodies is a glycoprotein (MW ~150,000 daltons) comprising four polypeptide chainstwo light
chains and two heavy chains, which are connected by disulfide bonds (Figure 1). Each light chain has a molecular weight of ~25,000 daltons
and is composed of two domains, one variable domain (VL) and one constant domain (CL). There are two types of light chains, lambda () and
kappa (). In humans, 60% of the light chains are , and 40% are , whereas in mice, 95% of the light chains are and only 5% are . A single
antibody molecule contains either light chains or light chains, but never both.
Each heavy chain has a molecular weight of ~50,000 daltons and consists of a constant and variable region. The heavy and light chains contain a
number of homologous sections consisting of similar but not identical groups of amino acid sequences. These homologous units consist of about
110 amino acids and are called immunoglobulin domains. The heavy chain contains one variable domain (VH) and either three or four constant
domains (CH1, CH2, CH3 and CH4, depending on the antibody class or isotype). The region between the CH1 and CH2 domains is called the hinge

region and permits flexibility between the two Fab arms of the Y-shaped antibody molecule, allowing them to open and close to accommodate
binding to two antigenic determinants separated by a fixed distance.
The heavy chain also serves to determine the functional activity of the antibody molecule. There are five antibody classesIgG, IgA, IgM, IgE
and IgDwhich are distinguished by their heavy chains , , , and , respectively (Table 1). The IgD, IgE and IgG antibody classes are each
made up of a single structural unit, whereas IgA antibodies may contain either one or two units and IgM antibodies consist of five disulfidelinked structural units. IgG antibodies are further divided into four subclasses (often referred to as isotypes) although the nomenclature differs
slightly depending on the species producing the antibody (Table 1).
Structure/function studies on IgG have been aided by the discovery that the proteolytic enzymes pepsin and papain cleave the molecule into
specific fragments with specific biological properties. Treatment of an IgG molecule with pepsin generates the F(ab')2 fragment, which broadly
encompasses the two Fab regions linked by the hinge region. Because the F(ab')2 molecule is bivalent, it is capable of precipitating an antigen.
Papain cleaves the IgG molecule in the hinge region between the CH1 and CH2 domains to yield two identical Fab fragments, which retain their
antigen-binding ability, and one non-antigenbinding fragmentthe (Fc) region. The Fc region is glycosylated and has many effector functions
(e.g., binding complement, binding to cell receptors on macrophages and monocytes), and serves to distinguish one class of antibody from
another.

Figure 1. Schematic representation of an antibody molecule.

Table 1. Overview of antibody classes and subclasses.

Antibody

IgA
IgE
IgD
IgM

IgG

Light
Chain
or
or
or
or
or

Light
Chain
or
or
or
or

Human and Mouse

Subtype
IgA1
IgA2

None
None
None

Human
Subtype
IgG1
IgG2
IgG3
IgG4

Heavy
Chain
1
2

Heavy
Chain
1
2
3
4

Light
Chain
or
or
or
or

Mouse
Subtype
IgG1
IgG2a
IgG2b
IgG3

Heavy
Chain
1
2a
2b
3

Antibody Structure and Classes of Immunoglobulins

This page introduces the nomenclature and criteria used to describe the structure, classes and functional
types of immunoglobulins.
Page Contents:

Structure of Immunoglobulins

Classes of Immunoglobulins
IgG IgM IgA IgD and IgE

Subclasses of Immunoglobulins

Polyclonal and Monoclonal Antibodies

Learn more...
Overview of Antibody
Production and Purification

Structure of Immunoglobulins
Antibody (or immunoglobulin) molecules are glycoproteins composed of one or more units, each
containing four polypeptide chains: two identical heavy chains (H) and two identical light chains (L). The
amino terminal ends of the polypeptide chains show considerable variation in amino acid composition and
are referred to as the variable (V) regions to distinguish them from the relatively constant (C) regions.
Each L chain consists of one variable domain VL and one constant domain CL. The H chains consist of a
variable domain, VH, and three constant domains CH1, CH2, CH3. Each heavy chain has about twice the
number of amino acids and molecular weight (~50,000) as each light chain (~25,000), resulting in a total
immunoglobulin monomer molecular weight of approximately 150,000.

Learn more...
Antibody Labeling and
Immobilization Sites

Related literature...

Antibody Production Handbook

Generalized structure of an immunoglobulin (IgG).

Heavy and light chains are held together by a combination of noncovalent interactions and covalent
interchain disulfide bonds, forming a bilaterally symmetric structure. The V regions of H and L chains
comprise the antigen-binding sites of the immunoglobulin (Ig) molecules. Each Ig monomer contains two
antigen-binding sites and is said to be bivalent.

The hinge region is the area of the H chains between the first and second C region domains and is held
together by disulfide bonds. This flexible hinge (found in IgG, IgA and IgD, but not IgM or IgE) region
allows the distance between the two antigen-binding sites to vary.

Classes of Immunoglobulins
The five primary classes of immunoglobulins are IgG, IgM, IgA, IgD and IgE. These are distinguished by
the type of heavy chain found in the molecule. IgG molecules have heavy chains known as gamma-chains;
IgMs have mu-chains; IgAs have alpha-chains; IgEs have epsilon-chains; and IgDs have delta-chains.
Differences in heavy chain polypeptides allow these immunoglobulins to function in different types of
immune responses and at particular stages of the immune response. The polypeptide protein sequences
responsible for these differences are found primarily in the Fc fragment. While there are five different
types of heavy chains, there are only two main types of light chains: kappa () and lambda ().
Antibody classes differ in valency as a result of different numbers of Y-like units (monomers) that join to
form the complete protein. For example, in humans, functioning IgM antibodies have five Y-shaped units
(pentamer) containing a total of ten light chains, ten heavy chains and ten antigen-binding sites (see
below).
IgG Immunoglobulins

IgG, a monomer, is the predominant Ig class present in human serum. Produced as part of the secondary
immune response to an antigen, this class of immunoglobulin constitutes approximately 75% of total
serum Ig. IgG is the only class of Ig that can cross the placenta in humans, and it is largely responsible for
protection of the newborn during the first months of life. Because of its relative abundance and excellent
specificity toward antigens, IgG is the principle antibody used in immunological research and clinical

View products...
IgG Purification Products
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diagnostics.
Properties of IgG:

Molecular weight: 150,000

H-chain type (MW): gamma (53,000)

Serum concentration: 10 to 16mg/mL

Percent of total immunoglobulin: 75%

Glycosylation (by weight): 3%

Distribution: intra- and extravascular

Function: secondary response

IgM Immunoglobulins

Serum IgM exists as a pentamer in mammals, predominates in primary immune responses to most
antigens, is the most efficient complement fixing immunoglobulin and comprises approximately 10% of
normal human serum Ig content. IgM is also expressed on the plasma mem- brane of the B lymphocytes as
a monomer. It is the B cell antigen receptor and the H chains each contain an additional hydrophobic
domain for anchoring in the membrane. Monomers of serum IgM are bound together by disulfide bonds
and a joining (J) chain.
Each of the five monomers is composed of two light chains (either kappa or lambda) and two heavy
chains. Unlike in IgG (and the generalized structure shown above), the heavy chain in IgM monomers is

Antibody Purification Methods

Antibody Fragmentation

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IgM Purification Kit

composed of one variable and four constant regions, the additional constant domain replacing the hinge
region. IgM can cause cell agglutination as a result of recognition of epitopes on invading
microorganisms. This antibody-antigen immune complex is then destroyed by
complement fixation or receptor mediated endocytosis by macrophages. IgM is
the first immunoglobulin class to be synthesized by the neonate and plays a role in
the pathogenesis of some autoimmune diseases.
Properties of IgM:

Molecular weight: 900,000

H-chain type (MW): mu (65,000)

Serum concentration: 0.5 to 2mg/mL

Percent of total immunoglobulin: 10%

Glycosylation (by weight): 12%

Distribution: mostly intravascular

Function: primary response

IgA Immunoglobulins

IgA exists in serum in both monomeric and dimeric forms, comprising approximately 15% of the total
serum Ig. Secretory IgA, a dimer, provides the primary defense mechanism against some local infections
because of its abundance in mucosal secretions (e.g., saliva, tears). The principal function of secretory IgA
may not be to destroy antigen but to prevent passage of foreign substances into the circulatory system.

View products...
Jacalin Agarose Human IgA
Purification Resin

Properties of IgA:

Molecular weight: 320,000 (secretory)

H-chain type (MW): alpha (55,000)

Serum concentration: 1 to 4mg/mL

Percent of total immunoglobulin: 15%

Glycosylation (by weight): 10%

Distribution: intravascular and secretions

Function: protect mucus membranes

IgD and IgE Immunoglobulins

IgD and IgE are found in serum in much smaller quantities than other Igs.
Membrane IgD is a receptor for antigen found mostly on mature B-lymphocytes.
IgE primarily defends against parasitic invasion and is responsible for allergic
reactions.
Properties of IgD:

Molecular weight: 180,000

H-chain type (MW): delta (70,000)

Learn more...
Antibody Isotyping Kits

Serum concentration: 0 to 0.4mg/mL

Percent of total immunoglobulin: 0.2%

Glycosylation (by weight): 13%

Distribution: lymphocyte surface

Function: unknown

Properties of IgE:

Molecular weight: 200,000

H-chain type (MW): epsilon (73,000)

Serum concentration: 10 to 400ng/mL

Percent of total immunoglobulin: 0.002%

Glycosylation (by weight): 12%

Distribution: basophils and mast cells in salive and nasal secretions

Function: protect against parasites

Subclasses of Immunoglobulins
In addition to the major immunoglobulin classes, several Ig subclasses exist in all members of a particular

Learn more...

animal species. Antibodies are classified into subclasses based on minor differences in the heavy chain
type of each Ig class.
In humans there are four subclasses of IgG: IgG1, IgG2, IgG3 and IgG4 (numbered in order of decreasing
concentration in serum). Variance among different subclasses is less than the variance among different
classes.

Mouse IgG Isotyping Kits

For example, IgG1 is more closely related to IgG2, 3 or 4 than to IgA, IgM, IgD or IgE. Consequently,
antibody-binding proteins (e.g., Protein A or Protein G) and most secondary antibodies used in
immunodetection methods cross-react with multiple subclasses but usually not multiple classes of Ig.

Polyclonal and Monoclonal Antibodies

Antibodies (whatever their class or subclass) are produced and purified in two basic forms for use as
reagents in immunoassays: polyclonal and monoclonal. Typically, the immunological response to an
antigen is heterogeneous, resulting in many different cell lines of B-lymphocytes (precursors of plasma
cells) producing antibodies to the same antigen. All of these cells originate from common stem cells, yet
each develops the individual capacity to make an antibody that recognizes a particular determinant
(epitope) on the same antigen. As a consequence of this heterogeneous response, serum from an
immunized animal will contain numerous antigen-specific antibody clones, potentially of several different
immuglobulin classes and subclasses comprising generally 2 to 5% of the total immunoglobulin. Because
it contains this heterogeneous collection of antigen-binding immunoglobulins, an antibody purified from
such a sample is called a polyclonal antibody. Polyclonal antibodies, which are generally purified directly
from serum, are especially useful as labeled secondary antibodies in immunoassays.
Because an individual B-lymphocyte produces and secretes only one specific antibody molecule, clones of
B-lymphocytes produce monoclonal antibodies. All antibodies secreted by a B-cell clone are identical,

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Antibody production
(immunogen preparation)