Introduction in Monoclonal antibodies

1. MONOCLONAL ANTIBODY
Saif Saleh Mohsen Ali
Molecular Immunology

2. Monoclonal & Polyclonal Antibodies
•Monoclonal antibodies (mAb):
• Are antibodies that are identical because they are produced by
one type of immune cell, all clones of a single parent cell.
•Polyclonal antibodies:
• Are antibodies that are derived from different cell lines. They
differ in amino acid sequence.

3. Difference Between Monoclonal and Polyclonal Antibodies

4. Difference Between Monoclonal and Polyclonal Antibodies

5. Practical steps in monoclonal antibody production:
1) Immunize animal .
2) Isolate spleen cells (containing antibody-producing B cells) .
3) Fuse spleen cells with myeloma cells (e.g. using PEG - polyethylene glycol) .
4) Allow unfused B cells to die .
5) Add aminopterin to culture to kill unfused myeloma cells.
6) Clone remaining cells (place 1 cell/well and allow each cell to grow into a clone
of cells) .
7) Screen supernatant of each clone for presence of the desired antibody .
8) Grow the chosen clone of cells in tissue culture indefinitely.
9) Harvest antibody from the culture supernatant.

6. Practical steps in monoclonal antibody production:

7. Monoclonal antibodies advantages
• 1) Homogeneity:
• Monoclonal antibody represents a single antibody molecule that binds to antigens
with the same affinity and promote the same
• effectors functions.
• 2) Specificity:
The product of a single hybridoma reacts with the same epitope on
• antigens.
• 3) Immunizing Antigen:
• Need not be pure or characterized and is ultimately not needed to produce large
quantities of antibody.
• 4) Selection:
• It is possible to select for specific epitope specificities and generate antibodies against
a wider range of antigenic determinants.
• 5) Antibody Production:
• Unlimited quantities of a single well-defined monospecific reagent.

8. Monoclonal antibodies disadvantages
• 1) Affinity:
• Average affinity of monoclonal antibodies are generally lower than polyclonal
antibodies.
• 2) Effector Functions:
Because antibody is monoclonal, it may not produce the
• desired biologic response.
• 3) Specificity:
• Monoclonal against conformational epitopes on native proteins may lose reactivity
with antigens.
• 4) Cross reactions:
• Antibodies sometimes display unexpected crossreactions with unrelated antigens.
• 5) Time and effort commitment: VERY LARGE.

9. Application of monoclonal antibodies
• Diagnostic Testing
•Pregnancy Testing
• Therapeutic uses
• Treatment of Cancer through Drugs
• Radioimmunodetection (RID) of Cancer
• Radioimmunotherapy (RIT) of Cancer
•Viral Disease Treatment
• Identifying Pathogens

10. Common Side effects of mAb
• Fever
• Chills
• Weakness
• Headache
• Nausea
• Vomiting
• Diarrhea
• Low blood pressure
•Allergies
•Cough
• Back pain
•General weakness
•Loss of appetite
•Insomnia
•Constipation
•Rashes