Monoclonal Antibodies for the diagnosis of diseases; the science, benefits and challenges.

1. UNIVERSITY FOR DEVELOPMENT STUDIES
FACULTY OF BIOSCIENCES
BTC 537: ANIMAL CELLAND TISSUE CULTURE
BY
ISSAH ABDUL-HANAN GIRASU
(M. PHIL BIOTECHNOLOGY)
EMAIL: abdul-hanan.issah14@uds.edu.gh

2. MONOCLONAL ANTIBODIES FOR DIAGNOSIS OF
DISEASES. THE SCIENCE BENEFITS AND CHALLENGES

3. What are Antibodies?
Antibodies are proteins produced and used by the immune system to identify and
neutralize foreign objects like bacteria and viruses.
These are also called immunoglobulins produced as a response to antigens.
Each antibody recognizes a specific antigen unique to its target.
There are 5 types of antibodies grouped based on their heavy chains; IgA, IgE, IgG,
IgD and IgM.

4. Structure of an Antibody

5. Classification
Antibodies can be classified into two; monoclonal and polyclonal by the means in
which they are created from lymphocytes.
Polyclonal antibodies (pAbs) are a mixture of heterogeneous usually produced by
different B cell clones in the body. They can recognize and bind to many different
epitopes of a single antigen. These are derived from different cell lines.
Monoclonal antibodies (mAb) are antibodies that are identical because they were
produced by one type of immune cell, all clones of a single parent cell.

6. Monoclonal Antibodies
Köhler and Milstein in 1995 discovered
antibody producing cells, by fusing murine
myeloma cells with murine-antibody-
secreting lymphocytes.
They found the hybridoma cells to have
single specificity, characteristic of
lymphocytes from immunized mice, but with
the myeloma cells’ ability to multiply
continuously.

7. Production of Monoclonal Antibodies

8. Preparation of Monoclonal Antibodies
The previous picture is showing a mouse being immunized against a
target cell “X” (antigens). This will allow the mouse to produce
antibodies for that will target against the “X” antigen.
Once the mouse has formed antibodies to the “X” antigen the
Antiserum (blood serum rich in antibodies) is extracted from the mouse’s
spleen.
 Monoclonal antibodies are produced by fusing single antibody forming cells
to tumour cells grown in culture. Results to hybridoma.
Hybridoma cells are continuously growing cell line generated by the
fusion of a myeloma cell and a normal cell that are capable of
producing antibodies.

9. Preparation of Monoclonal Antibodies Cont’d
Each hybridoma will produce relatively large quantities of identical antibody
molecules. Because the hybridoma is multiplying in culture, it is possible to produce
a population of cells, each is producing identical antibody molecules. These
antibodies are called "monoclonal antibodies" because they are produced by the
identical offspring of a single, cloned antibody producing cell.
HAT medium (Hypoxanthine Aminopetrin Thymidine) is used for preparation of
monoclonal antibodies, because it contains hypoxanthine, aminopterin, and
thymidine. This medium is selective for fused (hybridoma) cells. Unfused myeloma
cells and unfused spleen cells cannot grow because they lack HGPRT
(hypoxanthineguanine-phosphoribosyl transferase), and thus cannot replicate their
DNA and because of their limited life span respectively.

10. Diagnostic Applications of Monoclonal Antibodies
MAbs are utilized in diagnostic kits for the diagnosis of various infectious
diseases
For detecting pregnancies
For monitoring drug levels
For matching histocompatibility antigen,
For cancer and
In immunoscintography.
For diagnostic imaging to determine the extent of small cell lung cancer
(SCLC). By detecting tumour in different parts of the body at one time.

11. Diagnostic Applications of Monoclonal Antibodies
For example, the new agent, Nofetumomab, is a fragment of a monoclonal
antibody that when tagged with the radioisotope technique, can detect a
protein found on the surface of most small lung cancer cells.
Other monoclonal antibodies allows rapid diagnosis of hepatitis, influenza,
herpes, streptococcal, and Chlamydia infections.

12. Diagnostic Applications of Monoclonal Antibodies
• Monoclonal antibodies are used
to detect pregnancy in only 14 days
after conception.
• Urine and serum mAbs pregnancy
test are easier.

13. Diagnostic Applications of Monoclonal Antibodies
• Once monoclonal antibodies for a given
substance have been produced, they can
be used to detect the presence of this
substance. The Western blot test and
immuno dot blot tests detect the protein
on a membrane.

14. Diagnostic Applications of Monoclonal Antibodies
• mAbs are also very useful in immunohistochemistry, process of detecting
antigens (e.g., proteins) in cells of a tissue section by exploiting the principle
of antibodies binding specifically to antigens in biological tissues and
immuno fluorescence test, which detect the substance in a frozen tissue
section or in live cells.

15. MAJOR REFRENCE MATERIALS
• Fedorova VA, Samelia ZhG, Devdariani ZL, Utkin DV, Eremina OF, Liapina EP, et al. Development of
competitive immuno-assay based on monoclonal antibodies for the detection of specific antibodies to
pseudotuberculosis pathogen. Klin Lab Diagn. 2003;11:45–7. [PubMed] [Google Scholar]
• Marcilla A, Monteagudo C, Mormeneo S, Sentandreu R. Monoclonal antibody 3H8: a useful tool in the
diagnosis of candidiasis. Microbiol. 1999;145:695–701. [PubMed] [Google Scholar]
• Trowbridge IS. Interspecies spleen myeloma hybrid producing monoclonal antibodies against mouse
lymphocyte surface glycoprotein T 200. J Exp Med. 1978;148:313–23. [PMC free article] [PubMed] [Google
Scholar]

16. THANK YOU