Rapid SDS-PAGE Fluorescent Detection Method for Lactoferrin Content in Milk and Dairy Products
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Abstract:
A novel dye (Chromeo P503, P503) was used as an alternative to Coomassie brilliant blue to address the drawbacks associated with the laborious and time consuming (≥5 h) staining and destaining processes. This facilitated the establishment of a rapid and simple sodium dodecyl sulfonate-polyacrylamide gel electrophoresis (SDS-PAGE) method for the detection of lactoferrin in milk and dairy products. P503 dye-labeled protein emits strong red fluorescence, which could be completed in one step within 30 min. The interference of lactoferrin iron saturation on fluorescence signal was eliminated after apolactoferrin preparation to ensure the accuracy of quantification. Qualitative and quantitative detection of lactoferrin was achieved based on the presence of a 78 ku protein band and its gray value in the gel image. Under optimized experimental conditions, the linear range was 15~75 μg/mL, and the limit of detection was 5.11 μg/mL, with a recovery of 75.79%~108.09%, indicating that this method has high sensitivity and accuracy. The combination of the SDS-PAGE and P503 methods enabled the rapid, simple, and accurate detection of lactoferrin in milk and dairy products, eliminating the need pretreatment of the heparin affinity column. P503 dye labeling can facilitate rapid and low-cost quantitative detection of other milk proteins using SDS-PAGE, providing a valuable tool for dairy quality control, nutrition evaluation, and processing technology research.
