Monoclonal antibody generation

Our peptide antigen synthesis service is a fast efficient way of obtaining high quality peptide antigens. Using our in-house synthesis service, our peptide chemists can also assist with design of suitable peptide antigens and provide advice on any synthesis aspects if you wish to provide your own sequence for synthesis. Antigenic sections will be selected and the final peptide will be conjugated to a carrier protein to improve immunogenicity. Other possibilities are for the antigen to be internally sourced by AltaBioscience or supplied by the client.

We will immunise the antigen into a batch of 4 BALB/C mice, injecting initially three times over a period of 5 weeks and take serum samples on day 45 for assay. ELISA assays will be performed as part of service, using the immunised antigen. Serum samples (20-40ul) can also be despatched to the client for verification of specificity in further assays if required. If suitable titres are achieved, we will proceed to fusion. If poor responses are obtained, we will re-boost the animals (up to an additional 2 times) and take a further test sample(s).

No work can begin until Ethical Approval has been granted. Estimated turn around for this is 1-2 weeks from receipt of proposal.
We will only proceed to the fusion stage if satisfactory titres are achieved. If suitable titres are not achieved, the client has the option to terminate the programme at this stage, incurring no additional costs.

The ‘fusion’ cell culture stages include myeloma growth and preparation, fusion (using spleens from the two best responding mice), assay by ELISA of all colonies (20 x 96 well plates), cell culture maintenance and cryopreservation of up to 20 best binders.

For the assay, we will screen all colonies against the immunised antigen. Screening plates can be shipped to the client for further assays on the understanding that the testing is time sensitive.

In order to ensure that we have stable cell lines, candidates will require to be placed through rounds of limiting dilution cloning. We charge this on a per cell line basis of for each round of limiting dilution cloning (2-3 rounds of which might be required in order to ensure clonality).

ELISAs will be performed at each stage to assess the candidates, with results being provided at each stage. Plates can be provided for further testing by the client if required.

Upon confirmation of clonality and stability, lines will be scaled up and purified to produce 2-5mg of antibody (which will be shipped to the client). A working cell bank of 5 ampoules will also be created and stored for up to 1 year as part of service.

Additional Services

Cell lines can be scaled to any volume depending on specific requirements. Our processes use roller bottles cultures which allow for greater expression levels in comparison to other methods.

We can scale up cell lines into volumes of 500ml to 100 litres concurrently in either serum supplemented or serum-free media, with the scope of producing higher quantities in batches if required.

Serum-free media, in comparison to serum supplemented media, will ensure resultant purified mAb is homogeneous murine without any issue of contaminating with bovine antibody which could co-purify when using serum supplemented medium. Additionally, serum-free media tends to provide 25% greater antibody yield.