Custom Monoclonal Antibody Production
The APS Custom Monoclonal Project unit holds a licence under the
Animals (Scientific Procedures) Act. Our facilities are regulated and
controlled under strict UK Home Office guidelines ensuring a high
standard of animal care by experienced and dedicated staff and the
production of premier quality antibodies. Further information may
found on the Project requirements section of the INFORMATION page.
APS provides support at each stage of the Custom Monoclonal Project.
Project design support and regular technical discussions between the
customer and technical team ensure that that the project reaches a
successful outcome and a quality product is produced.
All APS Custom Antibody projects are treated in absolute confidence.
For Custom Monoclonal Projects all IP generated at any point of the
production of the hybridoma remains the property of the
Download APS Custom Monoclonal Antibody Production
The following four elements are considered before starting any project:
1. Choice of Antigen
2. Assessment of screening methods specifically in relation to the intended end use of the material
3. Hybridoma development and expectations
Custom Monoclonal Projects typically adhere to the following structure:
- Phase 1 – Antigen design and immunisation
- Phase 2 – Cell fusion and screening
- Phase 3 – Limiting dilution and cloning
- Phase 4 – Bulk antibody production
No project will continue to the next Phase without written confirmation being received from the client.
Antigen design and immunisation
For further information concerning the choice and selection
of a suitable antigen, please see our Antigen page or then contact our Technical team for further assistance.
Time: 6-10 weeks
Mouse Monoclonal Antibodies
The APS standard monoclonal antibody protocol involves using four female BALB/c mice, 6-8 weeks of age. These animals are immunised using our standard 81 day protocol, with immunisations being administered by either the sub-cutaneous or intraperitoneal route. A Project Schedule is provided giving expected immunisation and bleed dates. No project can be started without the written consent of the customer to this schedule.
Custom projects with additional immunisations and extended duration are designed following detailed discussion between the customer, the technical team and the animal unit.
Test bleeds are taken at regular intervals throughout the schedule and are sent to the customer for evaluation, unless a prior request is received for our technical team to evaluate the samples and provide the results.
Rat Monoclonal Antibodies
The APS standard rat monoclonal antibody protocol involves using two female Wistar rats, 8 weeks of age. The technical team are able to recommend a standard protoco,l but most customers prefer to work with the technical team to develop a protocol suitable for their requirements.
Screening and cell fusion
Time: 3-6 weeks
Custom monoclonal antibody projects allow for monitoring of the immunoreactivity of samples during the immunisation protocol. Once an immune response is detected further bleeds can be provided to the customer for additional confirmation. Once the immunisation protocol is complete the responses from the individual animals may be compared in order to select the most responsive donor animal for harvest of splenocytes prior to moving to the fusion step.
Once hyperimmune splenocytes have been identified, fusion with an immortalised cell line is performed using a Standard Operating Procedure. Initial screening by ELISA is carried out in order to identify the strongest candidate ‘binders’. There is opportunity at this stage to select isotype specific antibodies using defined secondary reagents.
Once cells expressing best antibodies have been identified they are expanded into 24-well plates for harvest of the tissue culture supernatant. This is required to enable further screening as previously determined using methods such as ELISA, Western Blot and FACS.
Antibody performance may then be assessed by the client – volumes of supernatant of approximately 5ml from the best colonies (up to 15) may be provided at this stage.
If, after five weeks in culture no hybridomas have been identified, the fusion is considered as unsuccessful. At this point the technical team will discuss best strategy to consider if a further fusion should be performed, or the project terminated.
Limiting dilution and cloning
Time: 3-5 weeks
Once the best colonies have been identified, limited dilution cloning is performed with the strongest parental colonies. Sub-clones with high specificity and expanded and 5 vials of each are cryopreserved. The resultant hybridoma cells and up to 5ml of supernatant from each sub-cloned cell line are then shipped to the customer.
Bulk antibody production
Time: 3-6 weeks
APS can offer further large scale production of hybridomas created as part of the Custom Monoclonal Antibody project, or from customer owned hybridomas.
Production from milligramme to gramme quantities is performed in vitro within bioreactor systems best suited to the specific hybridoma. APS offer a small scale pilot production service to evaluate existing hybridomas prior to offering quotes on large scale production. This allows our technical team to evaluate the hybridoma and to advise on optimisation prior to full scale production. Please see the Bulk Antibody Production section of our SERVICES page for further information.