Such hybrid cells are created through combining B-lymphocyte with tumour cells (myeloma cells).
These hybrid cell capable to yield monoclonal antibodies.
these cells produces monoclonal antibodies, and this technology is known as hybridoma technology.
In 1975, G. Kohler and C. Milstein discovered hybridoma technology and awarded to novel prize .
PRODUCTION
A hybridoma is created through the injection of a particular antigen into a mouse, obtaining the antigen-specific plasma cells (antibody-producing cell) from the mouse's spleen and
the succeeding fusion of this cell with myeloma cell.
Normal hybridoma technology was used for generation human mAbs from B lymphocytes from individuals infected with numerous pathogens or from cancer patients.
The technique has some components that are significant for the effective mAbs production.
1.Human B Lymphocytes
Unstimulated human B lymphocytes are scarcely utilized, they can be attached with human myeloma and yield mAbs.
2.Partner cells for fusion
There is plethora of myeloma cell fusion with human B cells.
Technique of Hybridoma Technology
firstly, B-lymphocytes are isolated from the animal spleen(mouse) which has been immunized with the required antigen against which monoclonal antibodies are produced.
Immunization of mouse is done by giving antigen injection alongside with an adjuvant .
Adjuvant is nonantigenic in nature however they stimulate the immune system.
This immunization with particular antigen surges the specific antibody making B-lymphocytes; this significantly surges the chances of attaining the vital hybridoma cells or clones.
This mixture of cell population is then cultured in selective media called as HAT (Hypoxanthine Aminopetrin Thymidine) medium along with the drug aminopterin.
Only the hybridoma cells have got the ability to divide and proliferate on the HAT medium.
Finally, select desired hybridoma for cloning and antibody production.Only one in several hundred cell hybrids will produce antibodies.
Culture selected hybridoma cells for the production of monoclonal antibodies in large quantities. Hybridoma cells can be frozen for future use.
PURIFICATION
Monoclonal antibodies may possibly require to be purified before they are expended for a range of applications.
Antibodies can be purified by following techniques
1.Ion-exchange chromatography
The method is powerful and can separate biomolecules that have minor differences in their
net charge. Eg. two protein molecules differing by a single charged amino acid.
2.Immobilized metal chelates chromatography Immobilized metal chelates chromatography (IMAC) usages chelate-immobilized divalent metal ions for the purification.
APPLICATION
1.Clinical importance in disease diagnosis
2.immunopurification
3.serological of ABO Blood groups.
