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Related Experiment Videos

Imaging surface plasmon resonance system for screening affinity ligands.

Paul R Morrill1, R B Millington, Christopher R Lowe

  • 1Institute of Biotechnology, University of Cambridge, Tennis Court Road, CB2 1QT, Cambridge, UK.

Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences
|August 9, 2003
PubMed
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A new imaging surface plasmon resonance (SPR) system accelerates ligand screening for affinity chromatography. This SPR system offers a faster, more efficient method for evaluating ligands for biopharmaceutical protein recovery.

Area of Science:

  • Biochemistry
  • Analytical Chemistry
  • Biotechnology

Background:

  • Affinity chromatography is crucial for biopharmaceutical protein purification.
  • Screening ligands for affinity chromatography is often time-consuming and resource-intensive.
  • Developing faster, more efficient screening methods is essential for bioprocess optimization.

Purpose of the Study:

  • To describe and evaluate a novel imaging surface plasmon resonance (SPR) system for screening affinity ligands.
  • To compare the performance of the SPR system with traditional chromatographic methods.
  • To establish a relational database correlating SPR screening data with chromatographic results.

Main Methods:

  • Synthesis and characterization of a combinatorial library of 13 ligands.

Related Experiment Videos

  • Immobilization of ligands onto agarose beads and gold SPR devices.
  • Measurement of binding and elution behavior against insulin analogue (MI3) and recombinant clotting factor (rFVIIa).
  • Fabrication of a durable, spatially addressable SPR transducer surface.
  • Utilizing a CCD-based imaging SPR instrument for real-time refractive index change detection.

    • Main Results:

    • The SPR system demonstrated a 15-fold increase in speed and required 130-fold less target protein compared to conventional methods.
    • Binding and elution data from SPR and chromatography were comparable, showing a strong linear correlation (R(2)=0.875) for K(AX) values.
    • The SPR transducer surface proved durable over >100 cycles and stable for 6 months.
    • The K(AX) values generated by SPR were approximately 2.5-fold higher than those from chromatography.

    Conclusions:

    • The developed imaging SPR system is a highly efficient and effective tool for screening and evaluating affinity ligands.
    • This SPR system provides a valuable alternative to traditional methods for ligand discovery in affinity chromatography.
    • The system shows significant potential for the recovery of biopharmaceutical proteins through optimized ligand selection.