Introduction to Hybridoma Technology
                     HYBRIDOMA TECHNOLOGY
WHAT IS 
HYBRIDOMA 
TECHNOLOGY?
Hybridoma technology is a well-established 
method to produce monoclonal antibodies 
specific to antigens of interest. 
Hybridoma cell lines are formed via fusion 
between a short-lived antibody-producing B 
cell and an immortal myeloma cell. 
Source: www.sinobiological.com
HYBRIDOMA
Each hybridoma constitutively expresses a 
large amount of one specific mAb, and favored 
hybridoma cell lines can be cryopreserved for 
long-lasting mAb production. 
As a result, researchers usually prefer 
generating hybridomas over other mAb 
production methods in order to maintain a 
convenient, never-ending supply of important 
mAbs.
Source: www.sinobiological.com
DISCOVERY
Hybridoma technology was discovered in 1975 
by two scientists, Georges Kohler and Cesar 
Milstein. They wanted to create immortal 
hybrid cells by fusing normal B cells from 
immunized mice with their myeloma cells.
Source: www.sinobiological.com
STEPS INVOLVED 
IN HYBRIDOMA 
TECHNOLOGY
Hybridoma technology is composed of several 
technical procedures, including antigen 
preparation, animal immunization, cell fusion, 
hybridoma screening and subcloning, as well as 
characterization and production of specific 
antibodies.
Source: www.sinobiological.com
GENERATION AND 
IDENTIFICATION
Generation and identification of high-quality 
hybridoma clones is a comprehensive and 
labor-intensive process, and requires months 
of work during the time frame from 
immunization to specific hybridoma 
identification.
Source: www.sinobiological.com
 CELL FUSION
Polyethylene glycol (PEG) and electrofusion are 
commonly used to induce cell fusion in 
hybridoma production. PEG fuses the plasma 
membranes of adjacent myeloma and/or 
antibody-secreting cells, forming a single cell 
with two or more nuclei.
Source: www.sinobiological.com
HYBRIDOMA 
SCREENING
Even in the most efficient hybridoma fusions, 
only about 1% of the starting cells are fused, 
and only about 1 in 105 form viable hybrids. 
This leaves a large number of unfused cells still 
in culture. 
Source: www.sinobiological.com
MAB 
PRODUCTION
Hybridoma antibodies can be produced in vitro 
and in vivo.
For production of monoclonal antibodies in 
vitro, hybridomas are expanded by transfer to 
24 well tissue culture plates followed by 25 
cm2 flask and a 75 cm2 flask containing 
suitable medium. 
Source: www.sinobiological.com
APPLICATIONS
The use of monoclonal antibodies is numerous 
and includes the prevention, diagnosis, and 
treatment of disease. For example, monoclonal 
antibodies can distinguish subsets of B cells 
and T cells, which is helpful in identifying 
different types of leukaemias. 
Source: en.wikipedia.org
BENEFITS OF 
HYBRIDOMA 
TECHNOLOGY
1. Natural affinity maturation
2. Low level of immunogenicity 
3. Constant domain activity
Source: en.wikipedia.org