Product Overview Data Sheet Protocol
pFB-CHIg-mG2ae2 is a cloning vector that expresses the mouse IgG2a heavy chain constant region with S239D/I332E mutations. It is a constitutive mammalian expression vector designed to deliver exceptionally high levels of antibody expression. This circular vector 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 vector carries an ampicillin resistance gene.
Characteristics
Fc engineered mouse IgG2a expression with S239D/I332E mutations:
– Increased binding to FcγRIIIa
– Increased ADCC
| Antibiotic Resistance | Ampicillin (AmpR) |
| Constitutive or Inducible System | Constitutive |
| Delivery Type | Transfection |
| Promoter | CMV |
| Product Type | Mammalian Expression Vector |
| Cloning Method | Restriction Enzyme (5’-AgeI; 3’-XhoI) or Homologous Assembly |
Contents & Storage
- 5.0 µg of pFB-CHIg-mG2ae2 in Tris-EDTA buffer
- Store at -20°. Vectors are guaranteed stable for 6 months when properly stored.
Materials required for Fc-engineered antibody generation
- pFB-CLIg-mk or pFB-CLIg-ml1 or pFB-CLIg-ml2 plasmid expressing the mouse kappa or lambda light chain.
Steps for Fc-engineered antibody generation
- Clone your heavy chain variable region (VH) into the pFB-CHIg-mG2ae1 vector to make a mouse heavy chain expression plasmid.
- Clone your light chain variable region (VL) into the pFB-CLIg-mk or pFB-CLIg-ml1 or pFB-CLIg-ml2 vector to make a mouse light chain expression plasmid.
- Co-transfect both heavy chain and light chain expression plasmids into your desired mammalian cell (such as CHO or HEK293) for Fc-engineered antibody production.
References
- Lazar et al., 2006. Engineered antibody Fc variants with enhanced effector function. Proc. Natl. Acad. Sci. USA 103, 4005–4010.
