Edible bird’s nest (EBN) hydrolysates have been proven to exhibit enhanced bioactivities. However, being a?macromolecule, fractions with different molecular weights would have different properties and bioactivities. Hence, this research was aimed to determine the chemical properties and?antioxidant activities?of freeze-dried (EBNFD) and spray-dried EBN (EBNSD) hydrolysates fractionated using?gel permeation chromatography?(GPC). Overall, two well-separated fractions were identified (EBNfFD1, EBNfFD2, EBNfSD1?and EBNfSD2). EBNFD?demonstrated significantly higher (P?≤ 0.05) peptide (3.6%), total carbohydrate (27.7%) and?sialic acid?(18.2%) contents than that of EBNSD. Similar trend was observed in low molecular weight fractionates (EBNfFD2?and EBNfSD2). Meanwhile, the first fractionates (EBNfFD1?and EBNfSD1) exhibited significantly higher (P?≤ 0.05)?hydroxyl radical?(?OH) scavenging activity. Fourier transform infrared (FTIR) spectroscopy demonstrated that all EBN fractionates have similar spectrum, except in the region of N—H (amide II) and C—H?alkyl group. In conclusion, EBN fractionates with different molecular weights showed different chemical properties and antioxidant activities.