Study on the Simultaneously Quantitative Detection for β-Lactoglobulin and Lactoferrin of Cow Milk by Using Protein...

Area of Science:

• Biotechnology
• Analytical Chemistry
• Immunosensing

Background:

• Accurate quantification of milk proteins like beta-Lactoglobulin (β-L) and Lactoferrin (Lf) is crucial for dairy quality control and infant nutrition.
• Existing methods for detecting β-L and Lf may not always allow for simultaneous, high-throughput analysis.

Purpose of the Study:

• To develop and validate a protein chip-based method for the simultaneous quantitative detection of β-Lactoglobulin (β-L) and Lactoferrin (Lf).

Main Methods:

• Utilized a protein chip printer to immobilize anti-β-L and anti-Lf antibodies.
• Employed an optimized sandwich assay format with specific detection antibodies for simultaneous antigen capture.
• Established standard curves on the protein chip and evaluated method precision and accuracy.

Main Results:

• Identified optimal antibody concentrations (0.5 mg/mL) and detection antibody titers (1:2,000) for both β-L and Lf.
• Achieved acceptable intra- and inter-assay variability (4.88%–38.33%).
• Demonstrated strong correlation with ELISA, with regression coefficients (r=0.734, P=0.038 for β-L; r=0.774, P=0.024 for Lf).

Conclusions:

• A robust protein chip method for simultaneous quantitative detection of β-Lactoglobulin and Lactoferrin has been successfully established.
• The developed method shows significant potential for further applications in protein analysis.