Quantitation of basic fibroblast growth factor by immunoassay using BIAcore 2000

Area of Science:

• Biochemistry
• Immunology
• Biosensor Technology

Background:

• Basic fibroblast growth factor (bFGF) is crucial in cellular processes.
• Accurate quantification of bFGF is essential for research and development.
• Existing methods for bFGF detection may lack sensitivity or efficiency.

Purpose of the Study:

• To develop and validate a sensitive, accurate, and efficient immunoassay for bFGF quantitation.
• To utilize a BIAcore 2000 biosensor instrument for bFGF detection.
• To establish a reliable method for bFGF analysis in various matrices.

Main Methods:

• Immobilization of anti-bFGF monoclonal antibody 48.1 (MAb 48.1) onto a Sensor Chip CM5 via amine coupling.
• Utilizing the immobilized MAb 48.1 sensor surface for bFGF detection and quantitation.
• Validation of the assay's sensitivity, accuracy, precision, and stability.

Main Results:

• The immunoassay demonstrated a reliable detection range of 5.65 to 1440 ng/ml bFGF.
• High immobilization level and reproducibility of MAb 48.1 were achieved.
• The assay showed excellent intra- and interassay precision (CVs < 8.5%) and stability over 150 regeneration cycles.
• No interference was observed from common biological and pharmaceutical matrices.

Conclusions:

• A validated, sensitive, and accurate immunoassay for bFGF quantitation using BIAcore technology has been developed.
• The assay exhibits high precision, accuracy, and specificity, making it suitable for research and development.
• The method's robustness and stability ensure its utility in diverse experimental settings.