pFUSE-CHIg-hG1e4

pFUSE-CHIg general plasmid map

Human IgG1 heavy chain constant region with increased ADCC and CDC

pFUSE-CHIg-hG1e4 is a cloning plasmid that expresses the human IgG1 heavy chain constant region with the E333A mutation. 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-hG1e4 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-hG1e4

• Encodes the human IgG1e4 CH constant region (E333A mutation)
• 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

References:

1. 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.
2. Idusogie EE. et al., 2001. Engineered antibodies with increased activity to recruit complement. J Immunol. 166(4):2571-5.

Specifications

• pFUSE-CHIg-hG1e4 features the constant region of the human IgG1 heavy chain with the E333A mutation

• pFUSEss-CHIg-hG1e4 features the constant region of the human IgG1 heavy chain with the E333A mutation and the IL2 signal sequence

Characteristics of E333A mutation:
- Increased binding to FcγRIIIa
- Increased 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