Product Overview Data Sheet Protocol
pFB-CHIg-hG1e7 is a cloning vector that expresses the human IgG1 heavy chain constant region with T307A/E380A/N434A 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 human IgG1 expression with T307A/E380A/N434A mutations:
– Increased binding to FcRn
– Increased half-life
| 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
- 20 µg of pFB-CHIg-hG1e7 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-hk or pFB-CLIg-hl plasmid expression the human kappa or lambda light chain.
Steps for Fc engineered antibody generation
- Cloning your heavy chain variable region (VH) into pFB-CHIg-hG1e7 vector to make heavy chain expression plasmid.
- Cloning your light chain variable region (VL) into pFB-CLIg-hk or pFB-CLIg-hl vector to make light chain expression plasmid.
- Co-transfecting both heavy chain and light chain expression plasmids into your desired mammalian cell (such as CHO, HEK293) for Fc engineered antibody production.
References
- Shields et al., 2001. High resolution mapping of the binding site on human IgG1 for Fc gamma, R.I.; Fc gamma RII, Fc gamma RIII, and FcRn and design of IgG1 variants with improved binding to the Fc gamma, R. J. Biol. Chem. 276, 6591–6604.
- Petkova et al., 2006. Enhanced half-life of genetically engineered human IgG1 antibodies in a humanized FcRn mouse model: Potential application in humorally mediated autoimmune disease. Int. Immunol.18, 1759–1769.
