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Antigens for Antibodies
Here you find information about antigens. What is an antigen, which antigens can you use to produce antibodies.

Monoclonal Antibodies

General Information

Monoclonal Antibodies

Monoclonal antibodies are produced from one B-cell. Every antibody is identical as they derive from one clone. Usually they only recognize one epitope. The development of monoclonal antibodies is more time consuming as you need to do different steps to generate good monoclonal antibodies.
Monoclonal Antibodies

Advantages of monoclonal antibodies

Monoclonal antibodies only recognize a single epitope. That means that you usually get less background and a clear signal in your applications. In addition you can produce a limitless amount of your antibodies as you receive the immortal antibody producing clone.

Production Steps

Monoclonal antibody production

In the following we describe the steps needed for monoclonal antibody production. At Davids Biotechnologie you can start and end with each step. I. e. you can order to immunize 2 x 5 mice and let us do the isolation of the spleen cells. You can continue with the fusion and screening by yourself or let us continue with the monoclonal antibody production.
Monoclonal Antibodies for further information.

STEP 1: Immunization

We immunize 5 mice, each with a different amount of antigen. So we ensure to get the optimal antigen concentration. We perform ELISA titer tests from each mouse and use the best mouse for further Steps.

STEP 2: Isolation of spleen cells

The accurate isolation of the spleen cells including the antibody producing B-cells is important to continue the monoclonal antibody production. The spleen cells can directly be used for fusion or they can be stored in liquid nitrogen. In the last case we can send you the cells for your own use.

STEP 3: Fusion

The isolated B-cells are fused with myeloma cells. Antibody producing hybridoma cells with endless growth are generated. The myeloma cells die in HAT-media. Only the hybridoma cells survive.

STEP 4: Screening

For the monoclonal antibody production the hybridoma cell with the best properties (recognition of the wanted epitope, growth, antibody production) has to be found. The screening is done with ELISA, DotBlot or IHC.

STEP 5: Subcloning

To ensure that the antibody clone is monoclonal (all antibodies come from one B-cell clone) we perform limited dilution.

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