TGEX-HC-hG1[LALA-PG]-Zeo

TGEX-HC-hG1[LALA-PG]-Zeo

Heavy Chain Cloning Vector for the Mammalian Expression of Human IgG1 Antibodies with an L234A, L235A, and P329G mutations in the CH2 region known to decrease ADCC and CDC in vitro.

Technical Description

The TGEX™ vector series is designed for the rapid expression of antibody molecules by transient gene expression in mammalian cell suspension cultures. TGEX™-HC-hG1[LALA-PG]-Zeo expresses the constant region of the human IgG1 heavy chain with an L234A, L235A, and P329G mutations in the CH2 region known to decrease ADCC and CDC in vitro (Schlothauer 2016). Expression of full length antibody molecules is achieved by co-transfection with a light chain variable region cloned into one of the TGEX™-LC vector series. Using transient transfection technologies yields of antibody between 20 mg/L and 200 mg/L and more in serum-free conditions can be obtained in just a few days.

Applications

• Mammalian expression of full length antibodies by transient gene expression.

• Selection of stable transfectants using Zeocin™.

For research use only; not intended for any animal or human therapeutic or diagnostic use.

Specifications

General Characteristics:

Plasmid Size: 5828
Promoter: CMV + TPL
Leader Peptide: pelBK
Cloning Site: Double BsaI
Fusion Protein: full length IgG1 human constant region with L234A, L235A, and P329G mutations
Selection: ampicillin (bacteria), zeocin™ (mammalian)
Replication: pMB1 high copy number

Physical Characteristics:

Concentration: 0.5 µg/µl.
Product Size: 10 µg.
Buffer: DNA Conservation Buffer (Tris/HCL 5 mM, EDTA 0.1 mM, pH 8.5, sterile).
Storage Temperature: -20°C.

Quality Control & Certification of Analysis

Product Sequence:

The entire plasmid sequence was confirmed by the Sanger method.

Certification:

Product meets all specifications.

References

1. Schlothauer, T. et al. 2016. Novel human IgG1 and IgG4 Fc-engineered antibodies with completely abolished immune effector functions. Protein Eng Des Sel, 29:457-466