A highly efficient electrofusion technique, vaccination protocols and ClonePix FL high throughput screening/selection procedures make up our hybridoma technology platform. It enables us to consistently deliver high quality mouse monoclonal antibodies in a high throughput, cost effective manner.
Our high throughput hybridoma platform:
- eliminates labour intensive / time consuming downstream screening and subcloning by limited dilution
- reduces the numbers of animals needed to generate hybridomas
- works for a broad range of antigens
- delivers fast results; 7-14 days to isolate selected clones
We guarantee to deliver ELISA antigen positive clones, using our standard protocols.
Typical Hybridoma Process, Deliverables and Time Lines
| Step | Process | Timeline |
|---|---|---|
| Immunization with clients antigen | Using 4 mice of the appropriate strain(s), we perform 4 vaccination rounds, each separated by 1-3 weeks. The first blood sample is taken 7 days after the 3rd injection. We screen the polyclonal serum samples by ELISA to determine comparative titres of antigen-specific antibody. The mouse with the best response receives a pre-fusion boost and is terminated for fusion. | 60-80 days |
| Electrofusion and selection | Direct fusion of mouse spleen lymphocytes with NS1 (or sp2/0) myeloma cells using MQR electrofusion protocol, and plating in 96-well plates. Hybridoma selection is performed by incubation in HAT media. | 1-2 weeks |
| Screening by ELISA | Within 10-14 days, surviving hybridomas as screened with an ELISA assay containing the antigen provided by the client. Screening is performed using secondary anti-mouse IgG. | 2 weeks |
| Expansion and cryopreservation | Up to 5 primary cultures are expanded for cryopreservation and single frozen copies of each are shipped to the client along with 1ml of the antibody-containing culture media from the cells. | 2 weeks |
| Subcloning of primary cultures | A maximum of 5 best primary cultures are subcloned 1x in semi solid media using our ClonePixFL automatic colony picking device. | 3-4 weeks |
| Expansion and cryopreservation | Each subcloned set is expanded and frozen into 3 vials at 2×106/ml, 1ml/vial. The client receives 2 vials of frozen cell and 4-10ml culture media the clones were growing in at the time they were frozen. 1 vial frozen cells is stored at MQR for 6 months as backup. | 2 weeks |
Watch The Video
Connect With Us
