Claims for Patent: 5,648,243
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Summary for Patent: 5,648,243
Title: | Human serum albumin expression construct |
Abstract: | The present invention provides DNA constructs comprising a promoter DNA sequence and a DNA sequence coding for human serum albumin. In one embodiment the human serum albumin sequence comprises at least one, but not all, of the introns in the naturally occurring gene encoding for the HSA protein. In another embodiment the DNA constructs comprise a 5\' regulatory sequence which directs the expression and secretion of HSA protein in the milk of a transgenic animal. Preferably, the promoter gene is a milk protein promoter sequence such as .beta.-lactoglobulin. The present invention also provides transgenic animals which secrete HSA in the milk of lactating females. The present invention also provides vectors comprising the constructs of the present invention. |
Inventor(s): | Hurwitz; David R. (Wyndmoor, PA), Nathan; Margret (West Chester, PA), Shani; Moshe (Mobile Post Modyin, IL) |
Assignee: | Rhone-Poulenc Rorer Pharmaceuticals, Inc. (Collegeville, PA) Peri Development Applications (1985) Ltd. (IL) |
Application Number: | 08/310,356 |
Patent Claims: | 1. A DNA construct adapted for the expression of human serum albumin in mammalian cells that do not normally express
human serum albumin, comprising a promoter from a mammalian gene and a sequence coding for human serum albumin,
(a) wherein the human serum albumin sequence comprises at least one, but not all, of the introns in the naturally occurring gene encoding the HSA protein; (b) wherein said introns are selected to provide for expression of HSA in said mammalian cells at levels equal to or greater than levels resulting from the expression of the naturally occurring HSA gene in said mammalian cells; and (c) wherein said naturally occurring HSA gene encodes the HSA protein and includes exons and introns in their native positional relationships. 2. The construct of claim 1 wherein said mammalian gene is a milk protein gene. 3. The genetic construct of claim 1 wherein said introns are selected from the group consisting of 1-6, 7-14, 1+7-14, 1+2+12-14, 2+7-14 and 1+2+7-14. 4. A DNA construct encoding HSA adapted for the expression of human serum albumin in mammalian cells that do not normally express human serum albumin, comprising one but not all of the first 7 introns of the HSA gene, and one of the last 7 introns of the HSA gene, (a) wherein said introns are selected to provide for expression of HSA in said mammalian cells at levels equal to or greater than the naturally occurring HSA gene in said mammalian cells; and (b) wherein said naturally occurring HSA gene encodes the HSA protein and includes exons and introns in their native positional relationships. 5. A DNA construct according to claim 1 comprising DNA sequences encoding human serum albumin operably linked to a mammary tissue specific promoter, said DNA construct expressed by the mammary glands of a lactating female transgenic mammal. 6. A DNA construct adapted for the expression of human serum albumin in mammalian cells that do not normally express human serum albumin comprising a sequence coding for human serum albumin and a promoter from a mammalian gene, said mammalian gene not encoding human serum albumin, (a) wherein the human serum albumin sequence comprises at least one, but not all, of the introns in the naturally occurring gene encoding the HSA protein, (b) wherein said introns are selected to provide for expression of HSA in said mammalian cells at levels equal to or greater than levels resulting from the expression of the naturally occurring HSA gene in said mammalian cells; and (c) wherein said naturally occurring HSA gene encodes the HSA protein and includes exons and introns in their native positional relationships. 7. A DNA construct comprising a promoter from a mammalian gene, and a sequence coding for human serum albumin, wherein the human serum albumin sequence contains an intron complement which is selected to provide expression of HSA in mammalian cells at levels greater than the levels of expression provided by a construct containing said promoter and the entire HSA gene with all 14 of its introns, in said mammalian cells. 8. A method of making human serum albumin in a mammalian cell that does not normally express human serum albumin comprising introducing the DNA construct of claim 1 into the mammalian cell, wherein human serum albumin is expressed in said mammalian cell at levels equal to or greater than levels resulting from the expression of the naturally occurring HSA gene in said mammalian cell. 9. The method of claim 8, further comprising the step of purifying the expressed human serum albumin. 10. A method of making human serum albumin in a mammalian cell that does not normally express human serum albumin comprising introducing the DNA construct of claim 2 into the mammalian cell, wherein human serum albumin is expressed in said mammalian cell at levels equal to or greater than levels resulting from the expression of the naturally occurring HSA gene in said mammalian cell. 11. The method of claim 10, further comprising the step of purifying the expressed human serum albumin. 12. A method of making human serum albumin in a mammalian cell that does not normally express human serum albumin comprising introducing the DNA construct of claim 3 into the mammalian cell, wherein human serum albumin is expressed in said mammalian cell at levels equal to or greater than levels resulting from the expression of the naturally occurring HSA gene in said mammalian cell. 13. The method of claim 12, further comprising the step of purifying the expressed human serum albumin. 14. A method of making human serum albumin in a mammalian cell that does not normally express human serum albumin comprising introducing the DNA construct of claim 7 into the mammalian cell, wherein human serum albumin is expressed in said mammalian cell at levels equal to or greater than levels resulting from the expression of the naturally occurring HSA gene in said mammalian cell. 15. The method of claim 14, further comprising the step of purifying the expressed human serum albumin. 16. A method of making human serum albumin in a mammalian cell that does not normally express human serum albumin comprising introducing the DNA construct of claim 5 into the mammalian cell, wherein human serum albumin is expressed in said mammalian cell at levels equal to or greater than levels resulting from the expression of the naturally occurring HSA gene in said mammalian cell. 17. The method of claim 16, further comprising the step of purifying the expressed human serum albumin. 18. A method of making human serum albumin in a mammalian cell that does not normally express human serum albumin comprising introducing the DNA construct of claim 6 into the mammalian cell, wherein human serum albumin is expressed in said mammalian cell at levels equal to or greater than levels resulting from the expression of the naturally occurring HSA gene in said mammalian cell. 19. The method of claim 18, further comprising the step of purifying the expressed human serum albumin. |
Details for Patent 5,648,243
Applicant | Tradename | Biologic Ingredient | Dosage Form | BLA | Approval Date | Patent No. | Expiredate |
---|---|---|---|---|---|---|---|
Merck Sharp & Dohme Corp. | INTRON A | interferon alfa-2b | For Injection | 103132 | 06/04/1986 | ⤷ Try a Trial | 2014-07-15 |
Merck Sharp & Dohme Corp. | INTRON A | interferon alfa-2b | For Injection | 103132 | ⤷ Try a Trial | 2014-07-15 | |
Merck Sharp & Dohme Corp. | INTRON A | interferon alfa-2b | Injection | 103132 | ⤷ Try a Trial | 2014-07-15 | |
>Applicant | >Tradename | >Biologic Ingredient | >Dosage Form | >BLA | >Approval Date | >Patent No. | >Expiredate |
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