ProMab Biotechnologies provides a high-quality hybridoma sequencing service, also called hybridoma development. Hybridoma sequencing specifically refers to the sequencing of the antibody variable regions of monoclonal antibodies being produced from hybridoma cell lines, and will be required for the production of humanized antibodies, or for the authentication of a hybridoma cell line.
Total RNA is extracted from 100,000 hybridoma cells, and using PCR methods, and the known non-variable flanking constant region sequences of respective antibody isotypes, the antibody variable regions (both heavy chain, VH, and light chain, VL) will be amplified for cloning and sequencing. Antigen-antibody binding will subsequently be confirmed by transient expression of either single-chain antibodies (scFv) or whole IgGs in a mammalian expression system. Further testing will be performed to confirm that the recombinant scFv or whole IgG retains antigen binding affinity and specificity to its target molecule.
Therefore, starting from a hybridoma cell line, we offer the following services:
• Validation of the hybridoma cell line in terms of antibody production, antibody isotyping and antigen-binding specificity;
• RNA extraction and reverse transcription;
• scFv sequencing by PCR amplification and subcloning of the variable domains;
• Full antibody sequencing by subcloning both variable and constant domains.
Contact us today to learn more about our hybridoma development services and how we can help you achieve your research goals.
Additional available services:
|Cloning and sequencing hybridoma antibody heavy and light chain variable regions||1 hybridoma||$1,400|
|Cloning and sequencing hybridoma antibody heavy and light chain variable regions + 100 ug production of reconstructed scFv-Fc||1 package||$3,800|
|Cloning and sequencing hybridoma antibody heavy and light chain variable regions + 100 ug production of reconstructed whole IgG||1 package||$3,800|
|Cloning and sequencing hybridoma antibody heavy and light chain variable regions + 100 ug production of reconstructed scFv-Fc + antigen-antibody binding confirmation using scFv||1 package||$6,600|
|Cloning and sequencing of hybridoma antibody + 100 ug production of reconstructed whole IgG + antigen-antibody binding confirmation using whole IgGs||1 package||$6,600|
1-2 weeks for cloning and sequencing antibody variable regions of hybridoma cell lines. An industry record: 4 days.
2-3 weeks for molecular construction, production and purification of IgG or scFv-Fc from mammalian cells, such as CHO and HEK 293.
1-2 weeks for assessment of the binding affinity and specificity of IgG or scFv.