1. Native and cleaved alpha 1-proteinase inhibitor was purified from ostrich serum using Sepharose-blue dextran chromatography, ammonium sulfate precipitation and ion exchange chromatography on DEAE-Toyopearl 650 M at pH 8.8 and 6.5. Ostrich alpha 1-PI displayed M(r) values of 68,100 using gradient PAGE and 66,200 using Ferguson plots. Isoelectric focusing of ostrich alpha 1-PI in the pH range 3-10 revealed pI values of 4.84 and 4.91, and in the pH range 4-6 the characteristic microheterogeneity observed for mammalian alpha 1-PIs was displayed. The presence of sialic acid, hexoses and hexosamines was detected using chemical methods, but were found in much lower quantities as compared to alpha 1-PIs of other species. Western blot analysis demonstrated a positive reaction between the native and cleaved ostrich alpha 1-PIs and the antibodies to the ostrich alpha 1-PIs raised in rabbits. No cross-reactivity was demonstrated by Western blot analysis between human alpha 1-PI and antibodies to ostrich alpha 1-PI. The inhibitory effect of alpha 1-PI on elastase and chymotrypsin was also investigated.