I am interested in generating a monoclonal antibody. Where do I start?

Please get in touch with the head of the Monoclonal Antibody Core Facility . We will need as many information of the antigen as possible, i.e.. sequence, structure, localisation within the cell, species, etc) and the intended applications you'd like to use the antibody, i.e. immunoblotting, IHC, IF, Flow Cytometry, or ELISA detection. We also need a project description that explains why the generation of new antibodies against your target of choice is important for your work.

What species will be used to generate my mAbs?

The host selected for immunization depends on the type of protein (intra- or extracellular) and the species from which the protein is derived. For generation of antibodies against extracellular mouse proteins, knock-out mice can be imported for immunization. Usually we immunize two wild-type rats and one mouse.

Do I have to validate the antibodies?

Yes. You should validate the antibody supernatants in the applications you'd like to use the antibody afterwards. The validation assy should be set up before the final boost.

Can I use the supernatants for assays that I have designed?

Yes. For most assays you can directly use the supernatant from stable monoclonal cell lines. We are adding NaN3 to the supernatant to avoid bacterial contamination. Affinity purification is required for antibody conjugation, for the use of antibodies as capture reagents and for in vivo applications.

Does the Core Facility help with the screening process?

Usually we perform a first FACS/ELISA screen to identify supernatants binding to the antigen. However, depending on the intended assays, we can help with capture, competition or Sandwich ELISA or cell screening in 96-well FACS analyses.

How can I find out about the progress of my project?

As you can see from the time line in ‘How we Work’, the generation of mAbs is a long and multi-step procedure.  Information on a project can be obtained at any time by contacting the Core Facility.

How do I get billed?

If you are a HMGU researcher, we will request your PSP account. For external partners, we will generate an offer.  Once the offer has been accepted, we will start the project. The production process is divided into three steps, which are billed independently. We will only charge the steps that have been initiated.

Do you guarantee that I will get an antibody for my project?

We have stong experience in generating monoclonal antibodies, providing every project its best chances for success. However, we cannot guarantee that your peptide or protein will lead to high-affinity antibodies. We have quality control measures in place that will help to increase the chances of success, software programs to analyse antigens and know-how on screening strategies, but there are always proteins that fail to produce an antibody despite every prediction.

Feel free to contact us if you have further questions.