"Quality derives from the design," Alpha Lifetech Inc. has a comprehensive R&D platform with unsurpassed technical expertise, by which we can offer our customers the full range of mouse hybridoma development services, including antigen design, animal immunization and hybridoma characteristics, to meet their requirements for each project.
Hybridoma Production - Mouse Hybridoma & Monoclonal Antibody Service
A general representation of the hybridoma method used to produce monoclonal antibodies.
Laboratory mice (Balb/c mouse) are immunized with a series of injections of the antigen, over the course of several weeks. Once splenocytes are isolated from the mammal's spleen, the B cells are fused with immortalized myeloma cells (NS0). The fusion of the B cells with myeloma cells can be done using electrofusion or PEG. Fused cells (B cells fused with myeloma cells) are incubated in HAT medium (hypoxanthine-aminopterin-thymidine medium) for roughly 2 weeks, since they have the hypoxanthine-guanine phosphoribosyltransferase (HGPRT) gene, making them non-sensitive and alive to the HAT medium. The incubated hybridoma cells are then diluted into multi-well plates to such an extent that each well contains only one cell. Since the antibodies in a well are produced by the same B cell, they will be directed towards the same epitope, and are thus monoclonal antibodies.
The next stage is a rapid primary screening process, which identifies and selects only those hybridomas that produce antibodies of appropriate specificity. Our experienced team provides high-throughput hybridoma screening by ELISA, FACS and other image-based binding methodologies (e.g., microbeads), as well as antibody biochemical characterization to ensure the identification of hybridoma candidates with high potency and to meet the special requirements of functional assays and animal studies.
After the separation of the mixed hybridoma cells, single hybridoma cell will be inoculated into larger tissue culture flask. This maintains the well-being of the hybridomas and provides enough cells for cryopreservation and supernatant for subsequent investigations. The culture supernatant can yield up to 60 µg/ml of monoclonal antibody. The Developmental Processes of Mouse Hybridoma Production.PDF
Program Features of Mouse Hybridoma & Monoclonal Antibody Service
-- Stable Hybridoma, High level of Antibody and Great Affinity
-- Serum free Medium
-- Exceptionally high cell fusion rates (1-10 fusions/1,000 B-cells) for creation of hybridomas
-- High-throughput analysis of MAb pools (up to 50,000 samples / assay)
-- Wide array of in-vitro and in-vivo bybridoma screening and characterization assays
-- SPF animal facility
The construction of stable cell lines is a laborious and time-consuming job, we hope to free customers from this tedious work to a more challenge work by using our relevant services. Experts in Alpha Lifetech Inc. have extensive experience in the field of cell culture and cell line generation. We have developed several perfect stable cell lines and also produced a large number of antibodies and proteins through stable cell lines. Our specific biosimilar cell lines and hybridoma cell lines are available in-house, and we are also pleased to provide custom strains to meet our clients' individual requirement.
If you have any questions, please feel free to contact us at any time.