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Last Updated: April 26, 2024

Claims for Patent: 7,049,406

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Summary for Patent: 7,049,406

Title:	Hemoglobin mutants with increased soluble expression and/or reduced nitric oxide scavenging
Abstract:	The invention relates to novel recombinant hemoglobins having reduced nitric oxide scavenging and/or increased high soluble expression. The invention further relates to methods of increasing the soluble expression of recombinant hemoglobin by adding exogenous hemin in molar excess of the heme binding sites of recombinant hemoglobin.
Inventor(s):	Weickert; Michael J (Belmont, CA), Glascock; Christopher B (Louisville, CO), Mathews; Antony J (Houston, TX), Lemon; Douglas D (Louisville, CO), Doherty; Daniel H (Boulder, CO), Olson; John S (Houston, TX)
Assignee:	Baxter Biotech Technology SARL (Neuchatel, CH) William Marsh Rice University (Houston, TX)
Application Number:	10/107,871
Patent Claims:	1. 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 .beta. 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. 2. The method of claim 1, further comprising adding excess hemin in step (b). 3. The method of claim 1, wherein said mutation is selected from the group consisting of D73I, D73M, D73E, D73T, D73Y, K82D, K82E, K82G, K82P, K82Q, K82S, K82N, N102A and N102V. 4. A method of obtaining high expression of soluble recombinant hemoglobin wherein at least one .beta. globin of the recombinant hemoglobin has at least one mutation or combination of mutations selected from the group consisting of D73Y, D73I, D73M, D73E, D73T, D73K, D73L, K82D, K82E, K82G, K82P, K82Q, K82S, K82N, N102A, N102V, K82D(Providence)+D73G(Tilburg), K82D(Providence)+D73Y (Vancouver), V67W+D73E, V67W+D73I, V67W+D73Y (Vancouver), V67W+D73E+K82D V67W+D73I+K82D(Providence), V67W+D73Y (Vancouver)+K82D(Providence), V67W+K82E, and V67W+K82G, comprising: (a) adding excess hemin to a culture of a host cell capable of expressing the soluble recombinant hemoglobin; (b) inducing the host cell to express the soluble recombinant hemoglobin; and (c) purifying the soluble recombinant hemoglobin. 5. The method of claim 4, wherein the recombinant hemoglobin comprises at least one beta globin mutation or a combination of beta globin mutations selected from the group consisting of: D73Y; D73I; D73M; D73E; D73T; K82D; K82E; K82G; K82P; K82Q; K82S; K82N; N102A; and N102V. 6. The method of claims 2 or 4, wherein at least a 3-fold molar excess of heme is added.

Details for Patent 7,049,406

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Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Recordati Rare Diseases, Inc.	PANHEMATIN	hemin for injection	For Injection	101246	07/20/1983	⤷ Try a Trial	2017-05-02
>Applicant	>Tradename	>Biologic Ingredient	>Dosage Form	>BLA	>Approval Date	>Patent No.	>Expiredate

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