A new spot quality control for protein macroarray based on immunological detection

• A visible means of spot quality control for protein macroarray has been developed.
• 1% (w/v) Ponceau S in spotting buffer was found to be optimal.
• The detection of food-borne pathogens based on the macroarray has been achieved.
• Properties of the multi-analytical protein macroarray are described.

Protein macroarray is a new, simple and multiple biochemistry detection system, in which the test spots are more than 1 mm diameter and results directly visible and instrumentation is not necessary. This technology, however, possesses recognized problems with spot quality and uniformity, issues that can limit its application. Previous methods have been developed for spot quality control, but they are complicated or require specific instrumentation. Therefore, we have developed a spot quality control buffer supplement with Ponceau S (SQCB-PS) as a direct and visible means of monitoring spot quality on nitrocellulose membrane prior to hybridization. In this report, 1% (w/v) Ponceau S and 10% (v/v) glycerol in spotting buffer were found to be optimal for spot quality control and food-borne pathogens multiple detection. Furthermore, the sensitivity and specificity of the protein macroarray were not compromised by spotting buffer with Ponceau S. Under optimal conditions, this visible spot quality control method makes the detection more reliable, and should enhance the wider use of the technique.

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