T4. Monoclonal Antibodies
T4. Monoclonal Antibodies
2. Monoclonal antibodies.
Monoclonal antibodies come from a homogenous cell population, where they are products of
1 clone of plasma cells derived from B-cells, resulting in identical copies of immunoglobulins
with equal primary structure, binding site specificity and effector functions. The origin can be
spontaneous in case of cancer growth of plasma cells, for example, or artificial in case of
lymphocyte hybridoma.
Some properties of these monoclonal antibodies are that they are chemically identical,
monospecific (against only 1 epitope), effective in lower concentrations, non-precipitating and
they can be prepared in unlimited amounts.
3. Antibody production.
Antibodies can be produced by exposing an
organism to an antigen, but we can also increase
this production by hybridizing plasma cells to
myeloma cells, creating hybridomas that can be
then separated by properties.
3.2. Selection.
Method based on enzyme defects are usually used in selection. This consist in inducing enzyme
defects in myeloma cells, so they won’t be able to grow in selection media but hybrid cells do
will be able, because they compensate the loss of enzyme from the other cell. We normally
produce nucleotide synthesis defects, most used are:
The selection medium use to be HAT (hypoxanthine, aminopterin and thymidine) and it’s
known that addition of insulin into HAT medium increases hybridoma formation. Hypoxanthine
and thymidine are XXXXXXXXXXXXXXXXXXXXX of the enzymes, and aminopterin is a molecule
that blocks nucleotide synthesis de novo, that is, from amino acid and saccharides.
With this technology we can avoid problems like biocompatibility, such us in use of mouse
antibodies in humans, because they have a short average life in serum, they don’t substitute
effector function and they can induce undesirable immune reactions or formation of human
anti-mouse antibodies.
Single chain antibodies. They are created by covalent bond between variable
fragments and various effector molecules.
Catalytic antibodies. They have an enzyme instead of FC domains or their parts.
Abzymes. They have catalytic site equivalent to the binding site and specifically bind a
haptene. Either this and the previous ones are effective catalysts of organic synthesis
reactions, so we can create antibodies, for example, against the substrate or an
intermediate product of the enzyme reaction.