TERMINOLOGY:

ANTIBODY:
An immunoglobulin molecule that has a specific amino acids sequence by virtue of
which it interacts only with the antigen that induces its synthesis in cells of lymphoid
series that is plasma cells.

ANTIGENS:
Substances that when introduced into the body stimulates the production of antibodies.
They may be toxins, bacteria, virus, parasite, foreign blood cells and cells of
transplanted organs.

EPITOPE:
The site or portion on an antigen that is recognized and bound by a particular antibody
by (Beta cells or cytotoxic T cells)

Myeloma
A tumor composed of cells found in bone marrow.

Adjuvant
A substance that non specifically enhance the immune response to an antigen.

Hybridoma
A clone of hybrid cells formed by fusion of normal lymphocytes with myeloma cells.
B.Lymphocytes + myeloma cells = Hybridoma cells
Monoclonal Antibodies;
Monoclonal antibodies are antibodies experimentally produced against a single type of
cell or substance.

These are specific to one epitope.

Polyclonal Antibodies
Antibodies derived from different clones of B cells due to presence of many types
of specific structures called as epitopes on the surface of a single antigen are
called polyclonal antibodies.

Different polyclonal antibodies specific to each epitope are produced and no. of
polyclonal antibodies depend upon the no. of epitopes.

PRINCIPLE OF MABS
The principle of Mabs is to produce an antibody that is specific in nature and
produced against a specific in nature and produced against a specific cell or
substance i-e antigens

As they are specific in nature so they bind to only one epitope and hence can be
used for various diagnostic & treatment purposes.

SYNTHESIS OF MABS
The key to synthesize Mabs is Myelomas that is cancerous tumors that arise
from the uncontrolled division of plasma cells

Basic steps for Mabs production:

1. A Myeloma cell is fused to an activaed (antibody secreting) B cells (Plasma cell) to

form a Hybrid cell called as Hybridoma.

2. Each Hybridoma is screened for the desired monoclonal antibody and cultured to
produce Hybridoma clones.
PRODUCTION PROCESS

a) Mouse is injected with antigen of interest against which Mabs are needed.
Injection stimulates mouses immune system to form antibody producing plasma
cells.
b) Plasma cells from mouses spleen are removed
c) Plasma Cells are mixed with myeloma cells from another mouse by forcing many
cells to fuse together Hybridomas are produed in 7-10 days.
Plasma cell if unfused with myelomas, can not survive and dies.
d) This fusion of plasma cells and myeloma in a tissue culture results in Hybridomas
cells which are immortal and which produce a single type of antibody i-e the
Monoclonal antibody.
e) Since the injected antigen may have several isotopes, different Hybrdoma
clusters way be producing different antibodies so individual and allowed to grow
in a culture.
F) Each Hybridoma cell can multiply indefinitely (i-e myeloma charaieestic)
producing nono clonal antibodies that are extracted from cell culture.

ADVANTAGES OF MABS
Pure

Uniform

Specific

Can be sensitized to probe for their antigenic targets.

Reproducible

Can be used as research tools

APPLICATION OF MABS

Widespread Application of MABs can be classified into five categories.

A. Diagnostic Utilities

B. Therapeutic usages

C. Immunopurification

D. Miscellaneous applications

A. Diagnostic Utilities
Diagnostic utilites are focussed for use of MABs to detect and identify the

Presence of a particular antigen or of antibodies specific to an antigen in a

sample. This detection is based on formation of antigen antibody complex (Ag-Ab

complex) MABs can detect upto picogram (10g) level

Examples;

1. for the precise and accurate classification of Blood groups in humans e.g.

2. For clear distinctive and decisive detection of pathogens involved in producing

specific dreadful diseases.

3. For tumor diagnosis: as most tumor contains a Marker antigen Known as

carcinoembryonic antigen (CEA). MABs can given following information about
tumors.

a. The nature of tumor cell type.

b. Malignant or benign kind of tumors

 c. The early instances of melastasis.

4. Effectively & accurately used for the detection of particular chromosomes of a

given species

B.Therapeutic usages;
1.Radioactivity to target tumor cells

MAB,s are used to d-90elivers radioactivity to the target tumor cells e.g 131I, 90Y(yitrium),
67 Cu, 212Pb etc

2.To lower tissue and Bone marrow explants rejections

In the allografts (i.e tussue transplanted from other individuals) tissue rejection occurs
once to T-cell activation T-cells specific MAB,s are aimed to eradicate T-cells from either
bone marrow or circulatory system CD3, CD4, CD8 are preferred targets for MABs.

3. Passive immunity against diseases

For passive immunity MAB,s are produced against a specific antigen and then
introduced into the individual.

4. Purification of antigens

MAB,s are used for purification of antigens that are specific to concerned pathogen.
These highly purified antigens are employed as vaccines e.g polio vaccine, cholera
vaccine, small pox vaccine etc.

C. Immuno purification;
Immuno purification refers to purification of antigens present in small quantum in the
form of a mixture alone with several kinds of other molecules.

Highly specific and critical interaction of a MAB,s with any antigen is employed for
immune purification e.g MAB,s have been frequently utilized in the isolation of m RNA
via encoding the particular protein entity to which MAB,s are eventually specific.
D. Miscellaneous applications
1. Drug delivery targeting
for this purpose magic bullets are used which are drug conjugated MAB
molecule. It has 2 advantages.
i. the desired drug is prevented from circulation in the body so small
dosage may be administered.
ii. Useful for anti-neoplastic agents e.g methotrexate, etc.

2. Autoimmune & immunodeficiency disease

in both autoimmune as were as immune deficiency diseases the prevalent helper
and suppressor so, T. cell subset ratio gets disturbed appreciably.so T-cells
deficiencies may be monitored accurately by use of MAB directed against specific
T cell immunogens.

3. Detection of surface molecules

MAB,s have been suillfully employed to probe the surface of immune competent
cells.
MAB,s have also been used in the mapping of actual prevailing disruption of the
membrane determinants e.g major histocompatibility complex (MHC) antigens.

4. Veterinary & plant diagnostics

- Foots & mouth disease in animals levels in humans i.e HCG (human choriono
gonadotropin) hormone
- plants viral diseases.