JoVE - Journal of Visualized Experiments
JoVE Visualize
Contact Us

Advancing therapeutic monoclonal antibody analysis: A sequential injection lab-on-valve approach

  • 0LAQV, REQUIMTE, Laboratory of Applied Chemistry, Department of Chemical Sciences, Faculty of Pharmacy, University of Porto, Porto 4050-213, Portugal.
Journal of Pharmaceutical and Biomedical Analysis +

|

July 15, 2025

|

July 15, 2025

View abstract on PubMed

Summary

This summary is machine-generated.

A new automated sequential injection lab-on-valve system with a spectrometer efficiently quantifies therapeutic monoclonal antibodies (mAbs). This high-throughput method offers accurate analysis for biopharmaceutical quality control and process monitoring.

Area Of Science

  • Analytical Chemistry
  • Biotechnology
  • Spectroscopy

Background

  • Automated flow-based techniques enhance analytical precision and efficiency.
  • Sequential Injection Lab-on-Valve (SI-LOV) systems provide miniaturized, resource-efficient high-throughput analysis.

Purpose Of The Study

  • To assess the feasibility of a fully automated SI-LOV system coupled with a miniaturized spectrometer for therapeutic monoclonal antibody (mAb) analysis.
  • To evaluate the system's accuracy, linearity, limit of detection, and qualitative assessment capabilities for six different mAbs.

Main Methods

  • Utilized a fully automated SI-LOV system integrated with a miniaturized spectrometer.
  • Analyzed six therapeutic mAbs, including Rituximab (RTX).
  • Employed second-order derivative spectroscopy and mass-based calibration for quantification and qualitative assessment.

Main Results

  • Achieved accurate quantification of mAbs, particularly RTX, with excellent linearity (R > 0.996) and a low limit of detection (0.09 mg/mL).
  • Qualitative assessment via derivative spectroscopy revealed characteristic peaks related to amino acids in mAbs.
  • Demonstrated good repeatability (1.8%) and reproducibility (2.3%) with rapid analysis time (approx. 110 s per determination).
  • Accurately estimated RTX concentration in a surrogate cell culture supernatant (>96%).

Conclusions

  • The automated SI-LOV system is a robust and effective platform for high-throughput mAb analysis.
  • The system shows significant potential for routine quality control, process monitoring, and biopharmaceutical workflows.
  • This technology offers a precise, efficient, and automated solution for therapeutic antibody assessment.

Keywords: