APS are able to offer a wide range of additional
custom services, offering a flexible and personal
We can custom produce product to your
strict specification to meet your time frame
with complete confidentiality, complete traceability
and our first class customer service.
Please contact us to discuss custom services
or any other requirements that you may have.
Antibodies frequently require labelling to enable them to be detected in immunological assay systems. APS is able to provide a full range of labelling services designed to render the antibody visible in the assay, yet not impair the binding properties.
Available labels include:
An immunogen is an antigen or any substance that may be specifically bound by components of the immune system.
The immunogenicity of the chosen antigen is of paramount importance to the success of any custom polyclonal antibody project. The characteristics of the antigen must be determined as this will impact the choice of host and protocol used to create the antibody and the eventual success of the project.
Whatever antigen is used there will probably be multiple epitopes against which antibodies could be generated.
When choosing epitopes for antibody production it is best to look for those with exposed hydrophilic regions as these will be easily accessible for antibody binding. Hydrophobic regions are usually hidden within the protein.
- Plant and yeast antigens
- Other antigens
- Antigen Preparation
Bulk Antibody Production
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 offers 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. The technology adopted by the technical team also allows antibodies to be produced at higher concentrations than with standard techniques.
Roller Bottle Technology
Culture using roller bottles results in a reliable cost-effective and consistent production. Batches of antibody to a volume of 100 litres are achievable, translating to over 5 gramme of purified monoclonal antibody in a short period of time.
This method is also readily adapted to the efficient production of antibody from multiple cell lines concurrently.
Appliflex Wavebag Bioreactor
APS also has the availability of a 5-20 litre Wavebag bioreactor for the production of larger multiple gram quantities. The Wavebag offers lower shear stress levels than in conventional stirred cell culture bioreactors, with oxygen transfer directly controlled by altering the rocking rate and the angle of the bioreactor bag.
APS offers cell storage with in dual liquid nitrogen repositories within different locations. Cells are duplicated between each repository. Cell banks may be maintained for the short term whilst antibody projects are ongoing, or long term storage solutions are available for cell banking.
For the majority of cell lines, when producing a standard unit of 1 litre culture volume, the Working Cell Bank (WCB) consists of 5 ampoules. This is included as part of the 1 litre batch production.
Master Cell Banks (MCBs) of 50-200 ampoules may also be developed, on request.
Cell Line Optimisation
Occasionally cell lines will become difficult ͛ and demonstrate poor growth. Our technical team has developed several proprietary methods for improving cell viability when these problems appear. Please contact our technical team to discuss any issues you may encounter with cell lines.
Occasionally cell lines may exhibit a degree of clonal instability which will result in a reduction in expression levels and, if allowed to continue, a total loss of secretion. Providing the original antigen is available for use in an ELISA assay it is possible to re-clone hybridoma lines to improve performance. If clonality is retrievable cells will be banked and then supplied to the customer. Please contact our technical team for further information.
Genotyping using PCR
We can offer a PCR genotyping service with rapid turnaround times (5-7 days), a high degree of accuracy and using robust methodology.
Please contact us for competitive pricing and further information.
APS provides high quality peptides at up to 95% purity. Standard peptides are produced at a purity of >85%, which is sufficient to produce an antigenic peptide suitable for antibody production. All peptides are supplied with HPLC profile and confirmation of molecular weight and purity measured by Mass Spectrometry. Further analysis is available on request. Peptides may also be supplied as labelled with fluorochromes, conjugated with carrier proteins. Other modifications include phosphorylation, methylation, acetylation, amidation, biotinylation and stable isotope labelling. Please contact us for further information on the full range of modifications that are available.
APS offers a peptide design service using BLAST search data. We can assist in determining the best peptide sequence to synthesise in order to assure the successful outcome of your antibody project. Please contact us for further details on this service.
Protein Free Media
Monoclonal antibodies for use in research are commonly developed in media supplemented with foetal bovine serum. For many applications this is the preferred method as the potential presence of bovine immunoglobulin in the finished, purified product is not an issue.
Drug Discovery and Clinical Diagnostic arenas have differing requirements and techniques have been developed to enable cells to be adapted to protein-free environments. This has often resulted in the requirement for the development of new monoclonal antibodies suitable for serum-free conditions or the adaptation of existing hybridomas to serum-free environments. APS has developed a custom service providing cell line adaptation and subsequent scale up using protein free media ensuring product purity and consistency.
The standard unit of production is a litre 1 litre batch , prepared from either a frozen ampoule or a growing culture of cells. Cells are expanded through T flask culture and used to inoculate 2 x 500ml roller bottles at a standard seeding density. The roller culture is then grown to extinction (cell viability <20%) and the confluent supernatant harvested and purified. This process typically has a 4 week lead time.
An option of a 5-20 litre wavebag bioreactor production (Appliflex system) is available, which can be useful in the production of larger (multiple gramme) quantities. Please see the Bulk Monoclonal Antibody Production section (above) for further information.
Our cell culture team has considerable experience working with an extensive cross section of hybridomas and other mammalian lines, ranging from poorly secreting recalcitrant lines produced in academic labs through to high performance lines producing antibodies for pre-clinical studies. Cell supernatants can be provided as an un-processed x10 concentrate or can be affinity purified in-house. Please see Purification and Modification of Antibodies for more information.
Purification and Modification of Antibodies
Purification of antibodies from either supernatant or serum involves the separation of the antibodies from other proteins. Whilst some assays do not require the use of purified antibodies, purification is often required to avoid unwanted interactions with other protein molecules and components.
APS provide a variety of purification options:
- Protein A,G and L for the purification of IgG
- Affinity purification
- Ammonium Sulphate Precipitation
- Dialysis and Ultrafiltration
- Ion Exchange Chromatography
- IgM Purification
- IgY Purification
Further processing and modification options:
- Endotoxin testing
- Protein modification – Antibodies may be modified by fragmentation with the enzymes papain to yield the Fab fragment, or pepsin to yield the F(ab’)2 fragment
Our antibodies can be purified, modified and presented in a form to suit you. For further details, see (PDF) or contact our technical team to discuss your specific requirements.