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High efficiency CHO cell display-based antibody maturation.

Ruiqi Luo1,2, Yun Zhao1, Yingjun Fan1,2

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We enhanced a CHO cell display method for antibody maturation using engineered activation-induced cytidine deaminase (AID). This significantly boosted mutation rates and types, improving antibody and protein maturation efficiency.

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Area of Science:

  • Biotechnology
  • Molecular Biology
  • Protein Engineering

Background:

  • Antibody maturation is crucial for developing therapeutic proteins.
  • Previous methods utilized activation-induced cytidine deaminase (AID) in CHO cells for antibody gene mutation.
  • Further optimization was needed to increase the efficiency of this system.

Purpose of the Study:

  • To improve the efficiency of a CHO cell display-based antibody maturation system.
  • To enhance mutation rates and diversity in antibody genes.
  • To optimize the engineering of AID and related genetic elements.

Main Methods:

  • Reengineering the activation-induced cytidine deaminase (AID) enzyme.
  • Optimizing the nucleic acid sequences of the target antibody and AID genes.
  • Refining the protocol for AID gene transfection into CHO cells.
  • Utilizing CHO cell proliferation for library generation.

Main Results:

  • Achieved a greater than 10-fold increase in antibody gene mutation rate.
  • Increased the diversity of mutation types observed in antibody genes.
  • Significantly improved the overall maturation efficiency for antibodies and other proteins.
  • Demonstrated a more robust and efficient antibody maturation platform.

Conclusions:

  • The reengineered AID system and optimized protocols substantially enhance antibody maturation efficiency.
  • This improved system offers a powerful tool for protein engineering and therapeutic antibody development.
  • The enhanced method provides greater mutation rates and diversity for antibody libraries.