Antibody Structure and Function

Antibodies

- are immunoglobulins, which react specifically with the antigen that stimulated their production

- make up about 20% of the plasma proteins.

Antibodies generated in response to a single complex antigen are heterogeneous because they
are formed by many different clones of cells.

Types of Antibodies:

Polyclonal - antibody capable of reacting with a different antigenic determinant on the complex
antigen; heterogenous

Monoclonal - antibody that arise from a single clone of cells, such as a plasma cell tumor
(myeloma); homogenous

 Monoclonal antibodies can be produced in vitro by fusing a myeloma cell with an

antibody producing B lymphocyte
Immunoglobulin (Ig) molecules share common structural features: all the Ig molecules are
composed of light and heavy polypeptide chains

The simplest antibody has a Y shape and consists of 4 polypeptide chains: 2 H chains and two L
chains. The 4 chains are covalently linked by disulfide bonds.

Light chains

 25,000 molecular weight

 can be either:

o κ (kappa)

o λ (lambda)

 can occur in all classes of immunoglobulins (IgG, IgM, IgA, IgD, and IgE)

 any one Ig molecule contains only one type of L chain.

 amino terminal portion of each L chain contains part of the antigen-binding site.

 L chain is composed of one variable domain (VL) and onconstant domain (CL)

Heavy chains

 50,000 molecular weight

 distinct for each of the five immunoglobulin classes and are designated:

o γ (gamma)

o μ (mu)

o α (alpha)
 o δ (delta)

o ε (epsilon)

 The amino terminal portion of each H chain participates in the antigen-binding site;

 the other (carboxyl) terminal forms the Fc fragment

 H chains have one variable domain (VH) and three or more constant domains (CH) Each
domain is approximately 110 amino acids in length.

Fc portion - participates in various biologic activities such as complement activation.

- is involved in placental transfer, complement fixation, attachment to various

cells, and other biologic activities.

Papain - a proteolytic enzyme that can split IgG into two fragments

Fab portion - fragment associated with antigen-binding activity when the peptide bonds in the
hinge region are broken.

The L and H chains of an Ig molecule are subdivided into:

a. variable regions - involved in antigen binding

b. constant regions - responsible for the biologic functions

The regions are composed domains

Domains - composed of three-dimensionally folded, repeating segments

High-resolution x-ray crystallography - used to determine structure of these domains

Hypervariable - are subregions consisting of extremely variable amino acid sequences form the
antigen-binding site

Epitope - antigenic determinant

Complementarity-determining region (CDR) - area where the the hypervariable regions form the
area of the Ig molecule complementary in structure to the antigenic determinant

Hybrid Virus - two viruses infecting a single host cell can swap genetic material, or reassort,
creating hybrid strains with characteristics of each parent virus

Cell Line Production - development requires the discovery of single cell-derived clones that
produce high and consistent levels of the target therapeutic protein. The first step in the
process is the isolation of single, viable cells. Limiting dilution is a technique that relies on
statistical probability but is time consuming.