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Hybridoma technology is used for the large-scale production of monoclonal antibodies. Monoclonal antibodies bind to only a single antigenic determinant or epitope. Such antibodies are used in research, diagnostics, and disease therapy. The hybridoma technology established in 1975 by Georges Köhler and Cesar Milstein was awarded the Nobel Prize in Medicine in 1984 for revolutionizing research and therapy.
Hybridoma Selection
Commonly used fusion techniques — electroporation,...
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Identifying PD-1/PD-L1 Inhibitors with Surface Plasmon Resonance Technology
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Non-affinity platform for processing knob-into-hole bispecific antibody.

Xiaoyang Wang1, Min Li1, Mengting Li1

  • 1Shanghai AsymBio Biotechnology Co., Ltd, Building 8, No.12, Lane 855, Jinzheng Road, Jinshan Industrial Park, Shanghai, China.

Bioresources and Bioprocessing
|December 18, 2024
PubMed
Summary
This summary is machine-generated.

This study introduces a novel, simplified purification method for bispecific antibodies, achieving high purity and recovery. The non-protein A platform offers a competitive alternative for biotherapeutic development.

Keywords:
Bispecific antibodiesByproductsHomodimerNon-protein A purification

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Area of Science:

  • Biochemistry
  • Biotechnology
  • Protein Purification

Background:

  • Bispecific antibodies offer therapeutic advantages but face development challenges due to byproduct formation.
  • High and low molecular weight variants, along with homodimers, complicate downstream purification.
  • Similar physicochemical properties of byproducts to target proteins hinder efficient separation.

Purpose of the Study:

  • To develop a non-protein A purification platform for bispecific antibodies.
  • To streamline purification processes and overcome challenges associated with byproduct removal.
  • To provide a competitive alternative to traditional affinity chromatography methods.

Main Methods:

  • A two-step chromatography approach was employed, utilizing mixed-mode Capto adhere resin for capture.
  • The capture step was performed at pH 7.90 ± 0.10.
  • Anion exchange chromatography served as the polishing step to achieve high purity.

Main Results:

  • The purification method yielded a final product purity of 98% by size-exclusion high-performance liquid chromatography (SEC-HPLC).
  • Reversed-phase-high-performance liquid chromatography (RP-HPLC) also confirmed 98% purity.
  • Residual host cell proteins were controlled at 10 ppm with an approximate 60% recovery rate.

Conclusions:

  • The developed non-protein A platform simplifies bispecific antibody purification.
  • This method offers a streamlined and cost-effective alternative to conventional protein A affinity chromatography.
  • The platform demonstrates significant potential for the commercial development of bispecific antibody therapeutics.