Chirality is an essential natural attribute of organisms. Chiral molecules exhibit differences in biochemical processes, pharmacodynamics, and toxicological properties, and their enantioselective recognition plays an important role in explaining life science processes and guiding drug design. Herein, we developed an ultra-sensitive enantiomer recognition platform based on an extended-gate metal-oxide semiconductor field-effect-transistor (Nafion–GO@BSA–EG-MOSFET) that achieved effective chiral resolution of ultra-sensitive Lysine (Lys) and α-Methylbenzylamine (α-Met) enantiodiscrimination at the femtomole level. Bovine serum albumin (BSA) was immobilized on the surface of graphene oxide (GO) through amide bond coupling to prepare the GO@BSA complex. GO@BSA was drop-cast on deposited Au surfaces with a Nafion solution to afford the extended-gate sensing unit. Effective recognition of chiral enantiomers of mandelic acid (MA), tartaric acid (TA), tryptophan (Trp), Lys and α-Met was realized. Moreover, the introduction of GO reduced non-specific adsorption, and the chiral resolution concentration of α-Met reached the level of picomole in a 5-fold diluted fetal bovine serum (FBS). Finally, the chiral recognition mechanism of the as-fabricated sensor was proposed.