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Related Experiment Video

Updated: Jan 14, 2026

Initial Evaluation of Antibody-conjugates Modified with Viral-derived Peptides for Increasing Cellular Accumulation and Improving Tumor Targeting
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3C conjugates: a highly sensitive platform for antibody internalization assessment in ADC development.

Changyan Chen1,2, Yutian Zhang1,2, Yadan Wu1,2

  • 1State Key Laboratory of Neurology and Oncology Drug Development, Simcere Pharmaceutical Group, Nanjing, China.

Mabs
|October 24, 2025
PubMed
Summary

A new 3C peptide conjugate platform enables sensitive, high-throughput antibody internalization analysis for antibody-drug conjugate (ADC) development. This versatile tool accelerates ADC discovery by providing reliable early-stage screening metrics.

Keywords:
3C peptideAntibody screeningAntibody-drug conjugationCytotoxicity assayInternalizationStreptococcal protein GpH-sensitive dye

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Oncology

Background:

  • Antibody-drug conjugates (ADCs) require antibody-mediated internalization for targeted payload delivery to tumor cells.
  • Accurate assessment of antibody internalization is crucial for efficient ADC development and antibody screening.
  • Conventional internalization assays present limitations in throughput, sensitivity, and specificity.

Purpose of the Study:

  • To introduce a novel 3C peptide conjugate platform for quantitative antibody internalization assessment.
  • To enable sensitive, high-throughput, and specific evaluation of antibody internalization for ADC discovery.
  • To provide a scalable solution for early-stage antibody screening in ADC development.

Main Methods:

  • Utilized streptococcal protein G's C1-C3 domains for high-affinity IgG binding.
  • Developed 3C-toxin for cytotoxicity-based internalization detection and 3C-pHAb for pH-sensitive fluorescent tracking.
  • Validated the platform across multiple tumor-associated targets (HER2, CDH6, LIV-1, LYPD3, GPC3).

Main Results:

  • Demonstrated sensitive and high-throughput antibody internalization evaluation without complex antibody modification.
  • Showed a strong correlation between 3C-based assays and the cytotoxic efficacy of corresponding ADCs.
  • 3C-toxin exhibited superior target promiscuity compared to traditional methods, broadening antigen applicability.

Conclusions:

  • The 3C peptide conjugate platform offers a versatile and scalable solution for antibody internalization analysis.
  • This platform accelerates ADC discovery by providing reliable early-stage screening metrics.
  • The developed assays overcome limitations of conventional methods, enhancing ADC development efficiency.