TGEX-HC-hG4[SPLE-PG]-Zeo

TGEX-HC-hG4[SPLE-PG]-Zeo

Heavy Chain Cloning Vector for the Mammalian Expression of Human IgG4 Antibodies with the S228P hinge stabilization mutation, and L235E/P329G mutations 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-hG4[SPLE-PG]-Zeo expresses the constant region of the human IgG4 heavy chain with the S228P hinge stabilization mutation, and L235E/P329G mutations 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 IgG4 human constant region with the S228P hinge stabilization mutation, and L235E/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