Product Overview Data Sheet Protocol
This vector is a constitutive mammalian expression plasmid designed to express high levels of human J-chain for assembly human pentamer IgM antibody expression with pFB-CHIg-hM (pentamer) and pFB-CLIg-hk or pFB-CLIg-hl. This circular expression plasmid features an enhanced, full-length CMV promoter and other expression elements that typically enable higher expression levels. It can be used in suspension-adapted cells, such as Expi293F™ and ExpiCHO™, for transient protein expression. Additionally, it can serve as a Geneticin®-selectable expression plasmid for engineering stable cell lines. The expression plasmid carries an ampicillin resistance gene.
Human IgM is the first antibody secreted by the adaptive immune system in response to a foreign antigen. Monomeric IgM is a heterotetramer of approximately 180 kDa. However, the secreted form of IgM exists predominantly in a pentameric configuration with a molecular weight greater than 900 kDa. The pentamer is connected at their Fc portions by a J, or joining chain (pFB-hIgJ).
Contents & Storage
- 20 µg of pFB-hIgJ in Tris-EDTA buffer
- Store at -20°. Plasmids are guaranteed stable for 6 months when properly stored.
Materials required for antibody generation and isotype switching
pFB-hIgJ, pFB-CHIg-hM (pentamer) with pFB-CLIg-hk or pFBCLIg-hl plasmid expressing the human kappa or lambda light chain.
Vector usage
- Entire monoclonal human IgM antibodies production: cloning your heavy/light chain variable regions in the cloning sites to preserve the integrity of the heavy/light chain constant region.
- Isotype switching: pFB-CHIg-hM (pentamer) with pFB-hIgJ and pFB-CLIg-hk or pFBCLIg-hl plasmids are designed to switch a monoclonal antibody from one isotype to another, therefore, maintaining the generated new antibodies with the same antigen affinity (epitopes) but with different effector functions (enhanced or reduced or even disabled ADCC and CDC).
