Despite the allergen labeling of peanuts, reducing peanut allergenicity through material selection and breeding may provide an avenue to reduce the severity of allergy due to consumption of foods containing peanuts. This study’s objective is to select peanut lines (cultivar/variety) with lower potent allergenic peptides. One hundred and twenty-two peanut lines (harvested in 2015, New Mexico State, USA.), including 97 lines from the Core USDA- Griffin collections were used for SDS-PAGE analysis in order to find low allergen peanut lines. Eleven peanut lines (Coded MS-1~MS-11) were identified as having lower allergen (Ara h 1, Ara h 2 and Ara h 3) levels and were re-planted in 2016 for further analysis. One line named MS-7 was selected for lower Ara h 1 (8.5-9.5% of total protein) and Ara h 2 (4.2-6.6% of total protein) content in 2015 and 2016. Two-dimensional (2-D) gel electrophoresis (isoelectric focusing plus SDS-PAGE) was conducted to compare the protein (allergens) distributions of MS-7 and check line (MS-9). The 2-D results supported the SDS-PAGE analysis that MS-7 had lower quantities of Ara h 1 and Ara h 2, the two major peanut allergens. The identified peanut line (MS-7) has potential for processing into lower allergenic peanut food products.