Edible bird’s nest (EBN)islong known as a delicacy and medicine among the Chinese communities all over the world. This work reports thephysicochemical, biochemical and antibacterial properties of EBN obtained from the swiftlet premises in Batu Pahat, Johor, Malaysia. Carbohydrates (46.47%) and proteins (35.8%) were the major constituents, with little fats (1.5%) and no fibre. Amino acid analysis by AccQ.Tag method revealed all eighteen types of amino acids including the eight essential ones. The heavy metal contents were within infant formula specification limits. Sodium, magnesium, potassium and calcium were the major minerals found. Alkaline conditions and heat treatment were the parameters that significantly affected the protein extraction from EBN. Out of the four EBN extracts (Run 1, Run 10, Oda and Goh) prepared; Run 10 had the highest protein content and anti-oxidant power, however, no direct correlations between protein content and the anti-oxidant power could be established. Oda and Goh extracts produced visible bands in the electrophoretic analysis in comparison to the Run 10 and Run 1 extracts. None of the extracts showed any antibacterial activity towards Escherichia coli, Klebsiella pneumoniae, Bacillus subtilis and Staphylococcus aureus.