NAME : NAIMA RAZA

ROLL NO : N - 60

THIRD PROFESSIONAL SECOND SEMESTER

PHARMACOLOGY ASSSIGNMENT

SUBMITTED TO : SIR UZAIR NASIR

ANTIBODIES :

Antibodies are large Y-shaped proteins. They are recruited by the immune system to identify and
neutralize foreign objects like bacteria and viruses.

Antibodies are sometimes used as a quick and potent immunosuppressive therapy to prevent the
acute rejection reactions as well as a targeted treatment of lymphoproliferative or autoimmune
disorders (e.g., anti-CD20 monoclonals).

MONOCLONAL ANTIBODY :

Monoclonal antibodies (mAb or moAb) are antibodies that are made by identical immune cells
that are all clones of a unique parent cell. Monoclonal antibodies can have monovalent affinity,
in that they bind to the same epitope (the part of an antigen that is recognized by the antibody).
Examples of therapeutic monoclonal antibodies

. Below are examples of clinically important monoclonal antibodies

Main
Type Application Mechanism/Target Mode
category

Anti- rheumatoid
inflamma arthritis
tory
Crohn's disease chimeri
infliximab inhibits TNF-
c
ulcerative colitis

ankylosing
spondylitis

rheumatoid
arthritis

Crohn's disease
adalimumab inhibits TNF- human

ulcerative colitis

ankylosing
spondylitis

inhibits IL-2 on chimeri

basiliximab acute rejection of
activated T cells c
kidney transplants

inhibits IL-2 on humani

daclizumab acute rejection of
activated T cells zed
kidney transplants
omalizumab inhibits humani
moderate-to- human immunoglobul zed
 severe in E(IgE)
allergic asthma
targets myeloid cell
surface humani
gemtuzumab relapsed acute
antigen CD33 on leuk zed
myeloid leukemia
emia cells
Anti- targets an
humani
cancer alemtuzumab antigen CD52 on T- a
B cell leukemia zed
nd B-lymphocytes

non-Hodgkin's
targets
lymphoma chimeri
rituximab phosphoprotein CD20
c
rheumatoid on B lymphocytes

arthritis

targets
breast cancer humani
trastuzumab the HER2/neu (erbB2
with HER2/neuover zed
) receptor
expression

approved
in squamous cell
carcinomas, Glioma humani
nimotuzumab EGFR inhibitor
zed
clinical trials for
other indications
underway
cetuximab EGFR inhibitor chimeri
approved c
in squamous cell
carcinomas, colorect
al carcinoma
 bevacizumab & ran humani
Anti-angiogenic inhibits VEGF
ibizumab zed
cancer therapy
Anti-
immunotherapy,
cancer chimeri
bavituximab cancer, hepatitis targets phosphatidylse
and anti- c
C infection rine[43]
viral

inhibits an RSV humani

palivizumab RSV infections
fusion (F) protein zed
in children
Other
inhibits the
prevent coagulati chimeri
abciximab receptor GpIIb/IIIa on
on in coronary c
platelets
angioplasty

USES :

Monoclonal antibody therapy is a form of immunotherapy that uses monoclonal

antibodies(mAb) to bind monospecifically to certain cells or proteins. This may then stimulate
the patient's immune system to attack those cells. ... It is possible to create a mAb specific to
almost any extracellular/ cell surface target.

WORKING OF MONOCLONAL ANTIBODIES HOW THEY ACT :

Monoclonal antibodies are designed to function in different ways. A particular drug may actually
function by more than one means. The role of the drug in helping the immune system may
include the following:

Flagging cancer cells. Some immune system cells depend on antibodies to locate the
target of an attack. Cancer cells that are coated in monoclonal antibodies may be more easily
detected and targeted for destruction.

Triggering cell-membrane destruction. Some monoclonal antibodies can trigger an

immune system response that can destroy the outer wall (membrane) of a cancer cell.
 Blocking cell growth. Some monoclonal antibodies block the connection between a
cancer cell and proteins that promote cell growth an activity that is necessary for tumor
growth and survival.

Preventing blood vessel growth. In order for a cancerous tumor to grow and survive, it
needs a blood supply. Some monoclonal antibody drugs block protein-cell interactions
necessary for the development of new blood vessels.

Blocking immune system inhibitors. Certain proteins that bind to immune system cells
are regulators that prevent overactivity of the system. Monoclonal antibodies that bind to
these immune system cells give the cancer-fighting cells an opportunity to work with less
inhibition.

Directly attacking cancer cells. Certain monoclonal antibodies may attack the cell more
directly, even though they were designed for another purpose. When some of these antibodies
attach to a cell, a series of events inside the cell may cause it to self-destruct.

Delivering radiation treatment. Because of a monoclonal antibody's ability to connect

with a cancer cell, the antibody can be engineered as a delivery vehicle for other treatments.
When a monoclonal antibody is attached to a small radioactive particle, it transports the
radiation treatment directly to cancer cells and may minimize the effect of radiation on
healthy cells. This variation of standard radiation therapy for cancer is called
radioimmunotherapy.

Delivering chemotherapy. Similarly, some monoclonal antibodies are attached to a

chemotherapeutic drug in order to deliver the treatment directly to the cancer cells while
avoiding healthy cells.

Binding cancer and immune cells. Some drugs combine two monoclonal antibodies,
one that attaches to a cancer cell and one that attaches to a specific immune system cell. This
connection may promote immune system attacks on the cancer cells.

SIDE EFFECTS OF MONOCLONAL ANTIBODIES :

 Allergic reactions, such as hives or itching.

Flu-like signs and symptoms, including chills, fatigue, fever, and muscle aches and pains.

Nausea, vomiting.

Diarrhea.

Skin rashes.

Low blood pressure.

POLYCLONAL ANTIBODIES :

Polyclonal antibodies (pAbs) are a complex mixture of several antibodies which are usually
produced by different B cell clones of an animal. These antibodies can recognize and bind to
many different epitopes of a single antigen and hence can form lattices with the antigens.

Production of Polyclonal Antibodies

PAbs are produced using a series of steps as described below:

Antigen Preparation
The quality of antigen used directly affects the immune response obtained. Even small amounts
of impurities will lead to antibodies reacting more to the impurity than to the desired antigen.
The quantity of the antigen is also crucial as it is proportional to the immune response evoked.
Too little or too much antigen may cause sensitization, suppression, or tolerance. Thus, the
antigen quantity should be determined based on several factors such as properties of the
particular antigen, the animal species chosen, injection frequency and the purity level of the
antigen.

Animal Species Selection

The animal species for the production of pAbs should be selected depending on factors such as:

the amount of PAb required,

the phylogenetic relationship between the animal and antigen,

the age of the animal,

the ease of obtaining blood samples, and

the application in which the PAb is going to be used.

Commonly used animal species in the lab for pAb production are the rabbit, rat, mouse, guinea
pig, hamster, goat, chicken, and sheep. Rabbits are more preferred due to their size and relatively
long life span.

Applications of Polyclonal Antibodies

Some key applications of PAbs are given below:

Pharmaceutical industry:

Digoxin Immune Fab is a pAb used to treat the fatal digitoxin toxicity.
 Rho(D) immune globulin is a pAb used to prevent a maternal immune response that
could transfer hemolytic disease to the newborn baby. The antibody binds passively to the
antigen and is produced from a pool of human plasma collected from Rhesus-negative
donors who have antibodies for the D antigen (present on red blood cells).

Histopathological analyses that use immunoperoxide staining

Affinity purification of antigens

Sandwich ELISA technique to detect tumor markers

Advantages of Polyclonal Antibodies

Some advantages of pAbs are listed below:

PAbs are a heterogeneous mix of antibodies that can bind to wide range of antigenic
epitopes. Hence, small change in the epitopes of an antigen is less likely to affect pAbs.

PAbs are very stable across a wide range of salt concentrations and pH values.

PAbs have a higher specificity compared to monoclonal antibodies (mAbs) as they have
specific antibodies for each epitope of an antigen.

PAbs are relatively easy to manufacture in a timely and cost-efficient way.

Disadvantages of Polyclonal Antibodies

Some disadvantages of pAbs are listed below:

 Unlike mAbs, pAbs produced against the same antigen in different animals will be
different in the case of immunized animals.

The affinity of pAbs to antigens may change over time.

The amount of pAbs produced is limited by the size and lifespan of the animal.

The purity and concentration levels of a specific antibody are lower in pAbs than that in
mAbs.

THERAPEUTIC USES :

Digoxin Immune Fab is the antigen binding fragment of polyclonal antibodies raised
to digitalis derivative as a hapten bound to a protein and is used for the reversal of life-
threatening digoxin or digitoxin toxicity.[

Rho(D) immune globulin is made from pooled human plasma provided by Rh-negative
donors with antibodies to the D antigen. It is used to provide passive immune binding of
antigen, preventing a maternal active immune response which could potentially result
in hemolytic disease of the newborn

POLYCLONAL ANTIBODY PRODUCTION :

Polyclonal antibody production will remain an essential research activity and rabbits will
continue to serve as one of the primary species used in polyclonal antibody production. The
schedules and methodologies used to immunize rabbits and produce polyclonal
antibodies will continue to vary dependent upon the immunogen, the adjuvant, and the end
purpose of the antibody. The choice of adjuvant and immunization schedules are critical
components of the polyclonal antibody production process that are frequently overlooked as
researchers resort to standard published methodologies (Cooper and Paterson, 2008, 2009;
Harlow, 1988) that may or may not be applicable to their needs. While FCA remains overall
the most effective adjuvant for polyclonal antibody production, the extensive histologic
 lesions it produces will continue to stimulate the development of alternative adjuvants
producing less tissue destruction

MECHANISM OF ACTION OF POLYCLONAL ANTIBODIES :

The exact mechanism of action is not fully understood, but polyclonal lymphocyte-depleting
antibodies work by binding to peripheral lymphocytes, blocking their function and targeting
them for destruction. Shortly after administration, these preparations lead to lymphocyte
depletion via complement-mediated lysis, removal by the reticuloendothelial system, and
antibody-dependent cell-mediated cytotoxicity (ADCC).18In contrast to other
immunosuppressive agents, polyclonal agents do not depend on T-cell activation but can
eliminate preactivated noncycling memory lymphocytes, which may be critical in presensitized
recipients. The more potent rabbit antithymocyte globulin Thymoglobulin has, for the most part,
replaced Atgam.
Dosing and Side Effects :
The usual dose of thymoglobulin is 1.5 to 2.0 mg/kg/day and 15 mg/kg/day for Atgam over 4 to
6 hours as an intravenous infusion. Thymoglobulin is recommended to be started during the
transplantation before reperfusion due to a decreased incidence of DGF rather than administered
postoperatively.19 The duration is usually 3 to 5 days for induction treatment and 5 to 10 days for
treatment of rejection. Polyclonal antibody preparations are foreign proteins and may cause a
variety of ad-verse effects. Most common are fever, chills, and arthralgias. Allergic reactions can
be avoided with premedication con-sisting of methylprednisolone, diphenhydramine, and
acetaminophen. Anaphylaxis and serum sickness occur rarely. Leuk-openia and
thrombocytopenia are common and need dose adjustment. Patients with white blood cell counts
between 2000 and 3000 or platelet counts between 50,000 and 75,000 require half-dose
Thymoglobulin, and it should be held if the white blood cell count is less than 2000 or the
platelet count is less than 50,000.18
HOW THEY USED FOR :

DIFFERENCES :

Polyclonal antibodies Monoclonal antibodies

Hybridomas take a relatively long time to
Relatively quick to produce
produce
Generate large amounts of non-specific Generate large amounts of specific
antibodies antibodies
Recognize multiple epitopes on any one
Recognize only one epitope on an antigen
antigen