Optimization of Peripheral Blood Mononuclear Cell Processing for Improved Clinical ELISpot Assay Performance

Area of Science:

• Immunology
• Biotechnology
• Clinical Research

Background:

• Cell and gene therapies show promise but face challenges from immune responses.
• Monitoring cellular immunogenicity is vital for long-lasting clinical efficacy.
• Enzyme-linked immunospot (ELISpot) assays are key for measuring immune responses but depend on peripheral blood mononuclear cell (PBMC) quality.

Purpose of the Study:

• To investigate the impact of blood sample processing time on ELISpot assay performance.
• To optimize sample processing conditions for robust cellular immune response assessment.
• To develop strategies to mitigate challenges in PBMC preparation for clinical studies.

Main Methods:

• Examined the effect of delayed blood sample processing on ELISpot assay signals.
• Optimized various sample processing parameters to recover ELISpot signals.
• Implemented mitigation strategies for granulocyte contamination in antigen-specific response detection.

Main Results:

• Overnight delays in blood sample processing led to a loss of ELISpot signals.
• Optimized processing successfully recovered ELISpot signals for samples processed within 32 hours.
• Mitigation strategies were identified to address granulocyte contamination issues.

Conclusions:

• Extended sample processing window for ELISpot assays in cell/gene therapy clinical studies.
• Provided solutions for logistical challenges in whole blood sample shipment and PBMC preparation.
• Enhanced the reliability of monitoring cellular immune responses in therapeutic development.