*Article* **Characterization and Extraction Influence Protein Profiling of Edible Bird's Nest**

**Nurul Nadiah Mohamad Nasir 1, Ramlah Mohamad Ibrahim 1, Md Zuki Abu Bakar 1,2, Rozi Mahmud 3 and Nor Asma Ab Razak 1,\***


**Abstract:** The edible bird nest (EBN) from *Aerodramus fuciphagus* has been consumed as a Chinese traditional food for health and medicinal purposes due to its elevated nutritional value. The present study focused on the influence of characterization and extraction methods on protein profiling, which could be a guideline for grading the EBN. The proposed extraction method is similar to the common food preparation methods of consumers and thus can accurately establish the bioactive protein available upon human consumption. The characterization includes physicochemical analysis (physical, morphology, elemental composition, and microbial content) and chemical analysis (crude protein and amino acid). The morphology of half-cup EBN was found to be uniformly shaped and rich in calcium as compared to rough surface of stripe-shaped EBN, and there was no significant microbial growth in both types of EBN. The crude protein and amino acid content in half-cup EBN were significantly higher than stripe-shaped EBN. The full stew (FS) and stew (SE) extraction methods produced a maximal yield of soluble protein. Sialic acid content in SE extract (8.47%, *w/w*) and FS extract (7.91%, *w/w*) were recorded. About seven parent proteins (39.15 to 181.68 kDa) were identified by LC-MS/MS Q-TOF, namely 78 kDa glucose-regulated protein, lysyl oxidase-3, Mucin-5AC-like, acidic mammalian chitinase-like, 45 kDa calcium-binding protein, nucleobindin-2, and ovoinhibitorlike. In conclusion, the characteristics and extraction methods influence the availability of bioactive protein and peptides, demonstrating the potential usage of EBN in improving its biological activities and nutritional properties.

**Keywords:** food; nutraceutical; edible bird's nest; *Aerodramus fuciphagus*; half-cup; stripe-shaped; physicochemical; proteomic; protein; extraction
