Patent ID: 7049406

Claim:
A method of obtaining high expression of soluble recombinant hemoglobin, comprising: (a) incorporating into providing a host cell a vector containing a gene nucleotide sequences encoding globins capable of associating to form a functional hemoglobin, wherein the nucleotide sequence encoding a β globin having at least one mutation or combination of mutations that directs high soluble expression of the recombinant hemoglobin selected from the group consisting of: D73Y: D73I; D73M; D73E; D73T; D73K; D73L; K82D; K82E; K82G; K82P; K82Q; K82S; K82N; K82H; N102A; N102V; K82D(Providence)+D73G(Tilburg); K82D(Providence)+D73Y(Vancouver); V67W+D73E; V67W+D73I; V67W+D73Y(Vancouver); V67W+D73E+K82D;(Providence) V67W+D73I+K82D;(Providence) V67W+D73Y(Vancouver)+K82D;(Providence); V67W+K82E; and V67W+K82G (b) inducing the host cell to express soluble recombinant hemoglobin; and (c) purifying the soluble recombinant hemoglobin.