Frequently Asked Questions
You can either fill out an online request form,
e-mail us at enquiries@oxfabs.com
or call us directly at +44 (0)1865 220911. We can then arrange to meet with you, or talk on the phone and discuss your specific project and customer needs. This means we can tailor a strategy to provide you with an accurate quote.
The production of an antibody requires an initial dialogue between the requesting customer and OxFabs to decide on the best approach to take. Once the strategy has been agreed the full service from molecule to monoclonal takes between 5 and 8 months.
The schedule below gives an idea of a minimum time scale for the production of an antibody.
| Timeline | Action | Approximate time taken |
| Day 1 | Customer completes an enquiry form | 1 day |
| Day 2 | Dialogue to establish project details Meeting set up with customer | 7 days |
| Day 9 | Customer receives quote | 7 days |
| Day 16 | Antigen preparation (recombinant protein) | 26.5 days |
| Day 43 | Immunisation starts | 33 days |
| Day 76 | Fusion performedHybridoma established | 13 days |
| Day 89 | Screening starts | 9-15 days |
| Day 104 | Cloning and freezing down | 28-46 days |
| Day 150 | Reagents given to customer | Total time = 5 months |
Yes, we provide an antigen design service. We consider this to be one of the most important contributors to the success of the project areas and it needs a full discussion before we start. In order for your antibody project to be as successful as possible, we can take a project from a gene name only and advise you on the best possible strategy for your project. We can recommend an antigenic peptide or the best regions of your molecule to use, in making a specific monoclonal antibody tailor-made to your exact needs. In depth discussion of the correct antigen design and screening strategy is the key to success.
We have developed antibodies in the past to a wide variety of antigens including recombinant proteins, synthetic peptides (including phosphopeptides) and cell preparations. This is not an exhaustive list and we are always pleased to work with any suggestions that may arise from the initial discussions we have. A detailed discussion at the beginning of the project will provide you with the relevant information to make an informed choice as to the best strategy.
Yes, OxFabs provides a peptide design service and can recommend and order your peptides conjugated to the appropriate carrier proteins for you. They will then be prepared appropriately in the correct buffer and stored ready for immunisation when required.
A minimum of 1mg should be sufficient for the immunisation and screening steps. Please contact us if you have less than this, and we can discuss the possibilities.
Antigen at a concentration of 1mg/ml is preferred but if your antigen differs from that please contact us to discuss.
We have already generated antibodies against insoluble proteins using inclusion bodies. We prefer to have a small amount of soluble protein for screening by ELISA. Please consult us if you have solubility concerns and our scientists will be more than happy to discuss and provide help.
Yes, we have a broad knowledge when it comes to immunisation, choice of myeloma strain and screening strategies. We can help you determine the best strategy for your project. Since antigen preparation can take some time, consult us before you start your project. Talk to us and let us help to develop your best monoclonal antibody generation strategy. One of our strengths is the comprehensive discussion that you will have with our scientists before and during antibody development. We believe that the initial planning with us is the reason why our antibodies have been so successful.
All of our antibodies are produced in vitro. The table below shows the steps involved, and gives an estimation of the time involved for each step and in total.
| Day 1 | First immunisation |
| Day 11 | Second immunisation |
| Day 21 | Third immunisation |
| Day 28 | Test bleed to check reactivity of serum |
| Day 31-33 | Final immunisation |
| Day 33-36 | Spleen cells harvested and fusion performed |
| Day 40-54 | Beginning of primary screening (usually by ELISA) of hybridoma colonies for antibody production |
| Day 50-54 | End of primary screening - at the end of this period it should be possible to identify whether the fusion has yielded any potential antibodies of interest |
| Day 40-54 | Growth of individual hybridoma colonies producing possibly potentially relevant antibodies and re-screening for reactivity with the target antigen |
| Day 45-60 | Cloning of hybridoma cell lines secreting specific monoclonal antibodies and storage in liquid nitrogen |
| Day 60-75 | Culture of clone(s) to produce large volumes of antibody (in the form of supernatant) for further characterisation |
Our first screening procedure will, in most instances, be by ELISA against the immunising antigen. Additional screening methods such as staining transfectants, immunohistological staining on normal and neoplastic tissues and western blotting are available. We have a broad range of experience in characterising monoclonals and we can advise you on the best approach. Please enquire regarding your specific project needs.
Evidence for the specificity of an antibody is normally provided by each of the following, where appropriate:
- Positive ELISA result against the immunising antigen.
- Positive labelling of transfected cells.
- Western blotting, showing reactivity with protein of the expected size.
- Immunohistological labelling of tissue sections, showing the expected or predicted cellular expression pattern.
The nature of biological processes is such that OxFabs cannot guarantee in each and every case to produce a satisfactory antibody, as there is an element of chance in all hybridoma fusions. To address this we have structured our payment system to reflect this and Terms and Conditions will be agreed at the time of Contract. However, we have had more than a 95% success rate in developing antibodies that will recognise the immunising antigen.
The hybridoma cell lines and the antibodies they produce are exclusively the property of the customer, unless other arrangements are made at the time of contract. We are happy to work jointly with the customer and consider other arrangements, such as sharing intellectual property (IP) which will have an input into the final project cost.
Yes, we can arrange and sign a confidentiality disclosure agreement (CDA) with the customer, if required, effective from the initial meeting.
Yes, we have over 30 years of experience in dealing with scientists from all over the world. If a face to face meeting is not feasible then the planning discussions can be achieved by other means - talking by phone and via e-mail, followed up by completing a comprehensive questionnaire is usually sufficient.
Services we can provide are:
- Monoclonal antibody development
- Antigen design and production
- Antibody production and purification
- Antibody characterisation
- Immunohistochemistry staining of normal and neoplastic tissues or cell lines
- Research and development
- Fluorescent in situ hybridisation (FISH)
- FISH and immunocytochemistry combined
- Workshops and teaching
If the services you require are not mentioned in the list shown above, please don't hesitate to contact us to discuss your requirements further. We have an extensive Skills Matrix available for viewing on request.