Three recombinant soybean cysteine proteinase inhibitors (rSCPIs) L1, R1, and N2 were chemically characterized. These inhibitors have the potential to inhibit the growth and development of three major agricultural crop pests known to utilize cysteine proteinases (CPs) for protein digestion: Western corn rootworm, Colorado potato beetle, and cowpea weevil. Characterization data obtained show differences between the inhibitors and will be needed to consider the use of rSCPIs to create insect resistance in plants.