Advantages of the Lab-on-a-Chip Method in the Determination of the Kunitz Trypsin Inhibitor in Soybean Varieties

Abstract

Qualitative and quantitative determination of the Kunitz trypsin inhibitor (KTI) in soybean experimental lines is very important in processes of selecting and breeding of new varieties. The total enzyme activity assay, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and Lab-on-a-Chip (LoaC) method were used for the determination of the presence and quantity of KTI in 15 soybean experimental lines and varieties. From the total trypsin inhibitor enzyme assay, inhibitor activities were registered in all samples, even in a Kunitz variety that was a negative control. The SDS-PAGE method did not detect the presence of the KTI protein band in seven soybean experimental lines and Kunitz variety, while the LoaC method showed the absence of KTI only in the Kunitz variety sample. Results confirmed the superiority of the LoaC method over other two methods in selectivity and sensitivity when KTI determination is concerned. Relationships between the KTI content obtained by the LoaC method and total trypsin inhibitor enzyme activity were established and statistically confirmed.