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The development of functional food ingredients using legume seed proteins has potential for nutraceutical use. We purified recombinant β-conglutin proteins (rβ1 to rβ4, and rβ6) from narrow-leafed lupin using affinity–chromatography, and evaluated their anti-inflammatory activity using ex vivo and in vitro systems. rβ1, rβ3, and rβ6 produced lower levels of pro-inflammatory mediators such as nitric oxide (about –34.0–fold in all cases), iNOS mRNA (−7.15, −7.97, −7.41–fold), interleukin 1β (−12.05, −11.64, −12.16–fold), chemokine CCL5 (−16.0, −18.0, −19.0–fold), and cytokines including TNF-α, INF-γ, IL-1β, IL-2, IL-6, IL-8, and IL-12 (β1: −28, −100, −8, −2, −49, −45, −127–fold; β3: −22, −400, −9, −3, −33, −10, −2.54–fold; β6: −72, −122, −11, −3, −2000, −13, −338–fold).These results suggest that the β1, β3, and β6 conglutins have potential as functional food components in nutraceuticals and that can provide alternative therapies for the prevention and treatment of inflammatory-related diseases.