Velvet antlers (VA) sourced from red (Cervus elaphus), sika (C. nippon), and elk (C. canadensis) deer have become increasingly popular in Chinese, Korean and Japanese traditional and oriental medicines as supplements for disease prevention. Therefore, this study aimed to determine the chemical components (proximate composition, amino, uronic, and sialic acids) and to characterize the bacterial profile of VA extract. The yield and contents of uronic and sialic acids extracted from VA at 98-100℃ for 29-36 sec were determined to be 40.71%, 28.06 mg/g and 0.73 mg/g, respectively. Overall, proximate parameters were higher in extracts recovered at 100℃ compared with 90℃, and these parameters were: moisture (4.66%), crude protein (86.63%), crude fat (3.23%), crude ash (4.1%), and carbohydrate content (5.43%). Alanine was the most abundant among the identified amino acids. Analysis of 16S rRNA sequence identified 12 bacterial species, including seven gram-positive and five gram-negative bacteria. Among all bacteria, Chryseobacterium indologenes, Shigella flexneri ATCC 29903, Staphylococcus equorum, Staphylococcus xylosus, Staphylococcus succinus, and Vagococcus fluvialis had 100% identity in the VA extract. The morphology of C. indologene, S. flexneri ATCC 29903, and V. fluvialis was bacilli, whereas S. equorum, S. succinus, and S. xylosus were cocci. Key words:velvet antler, chemical composition, characterization, amino acid, bacteria