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Chitosan was modified by coupling with linolenic acid through the 1-ethyl-3-(3-dimethylaminopropyyl)carbodiimide-mediated reaction. The degree of substitution was measured by 1H NMR, and it was 1.8%, i.e., 1.8 linolenic acids group per 100 anhydroglucose units. The critical aggregation concentration (CAC) of the self-aggregate of hydrophobically modified chitosan was determined by measuring the fluorescence intensity of the pyrene as a fluorescent probe. The CAC value in phosphate-buffered saline (PBS) solution (pH 7.4) was 5 x 10(-2) mg/mL. The average particle size of self-aggregates of hydrophobically modified chitosan in PBS solution (pH 7.4) was 210.8 nm with a unimodal size distribution ranging from 100 to 500 nm. A transmission electron microscopy study showed that the formation of near spherical shape nanoparticles had enough structural integrity. The loading ability of hydrophobically modified chitosan (LA-chitosan) was investigated by using bovine serum albumin (BSA) as a model protein. Self-aggregated nanoparticles exhibited an increased loading capacity (19.85 +/- 0.04 to 37.57 +/- 0.25%) with an increasing concentration of BSA (0.1-0.5 mg/mL).