pFUSE-CHIg-hG1e3

pFUSE-CHIg general plasmid map

Human IgG1 heavy chain constant region with reduced ADCC and CDC

pFUSE-CHIg-hG1e3 is a cloning plasmid that expresses the human IgG1 heavy chain constant region with E233P / L234V / L235A / deltaG236 + A327G / A330S / P331S mutations. It contains a multiple cloning site upstream of the constant region to enable cloning of the heavy chain variable region. This plasmid is designed for antibody generation when co-transfected into mammalian cells with the light chain variable region cloned into pFUSE-CLIg.

pFUSEss-CHIg-hG1e3 additionally features the IL2 signal sequence to allow the secretion of recombinant antibodies generated using fragment antigen-binding (Fab) or single-chain variable fragment (scFv)  fragments selected from phage display libraries that lack a signal sequence.

Key features of pFUSE-CHIg-hG1e3

• Encodes the human IgG1e3 CH constant region (E233P / L234V / L235A / deltaG236 + A327G / A330S / P331S mutations)
• Long-lasting expression due to the composite hEF1-HTLV promoter
• Selectable with Zeocin™ in both bacterial and mammalian cells
• Available with or without the IL-2 signal sequence

DISCLAIMER: pFUSE-CHIg-hG1e3 plasmids featuring the IgG1e3 engineered Fc are sold by InvivoGen under a limited license from Cambridge Enterprise Limited (International Publication Number WO 99/58572).

References:

1. Armour KL. et al., 1999.  Recombinant human IgG molecules lacking Fcgamma receptor I binding and monocyte triggering activities. Eur J Immunol. 29(8):2613-24.
2. Shields RL. et al., 2001. High-resolution mapping of the binding site on human IgG1 for Fc gamma RI, 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(9):6591-604.

Specifications

• pFUSE-CHIg-hG1e3 features the constant region of the human IgG1 heavy chain with E233P / L234V / L235A / ΔG236 + A327G / A330S / P331S mutations

• pFUSEss-CHIg-hG1e1 features the constant region of the human IgG1 heavy chain with E233P / L234V / L235A / ΔG236 + A327G / A330S / P331S mutations and the IL2 signal sequence

Characteristics of E233P / L234V / L235A / ΔG236 + A327G / A330S / P331S mutations:
- Reduced binding to FcγRI
- Reduced ADCC and CDC

Both plasmids are selectable with Zeocin™.

pFUSE Contents

• 20 µg of lyophilized DNA
• 1 ml of Zeocin® (100 mg/ml)

Product is shipped at room temperature