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Last Updated: September 22, 2021

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Claims for Patent: 5,739,407

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Summary for Patent: 5,739,407
Title: Human .beta.-casein, process for producing it and use thereof
Abstract:The present invention relates to a DNA sequence encoding the human milk protein .beta.-casein or an analogue or variant thereof which has either the calcium binding activity of human .beta.-casein, or opioid activity, or angiotensin converting enzyme (ACE) inhibitory activity, or a combination of any two or three of these activities. The DNA sequence may optionally contain one or more intron sequences and permissive RNA splice signals. The DNA sequence is used in the production of recombinant human .beta.-casein, advantageously by means of production in transgenic non-human mammals such as bovine species. In one embodiment, the DNA sequence is inserted into a milk protein gene of a mammal such as a whey acidic protein (WAP) gene. The main use of the recombinant human .beta.-casein is as a constituent of infant formulae. It is contemplated that the recombinant human .beta.-casein provides a substantial improvement of the nutritional and biological value of the formulae in that a closer similarity to human milk is obtained.
Inventor(s): Bergstrom; Sven (Umea, SE), Hernell; Olle (Umea, SE), Lonnerdal; Bo (Davis, CA), Hjalmarsson; Karin (Umea, SE), Hanson; Lennart (Umea, SE), Tornell; Jan (Goteborg, SE), Stromqvigt; Mats (Umea, SE)
Assignee: Symbicom aktiebolag (Umea, SE)
Application Number:08/078,090
Patent Claims:1. An expression system comprising a DNA sequence which encodes a polypeptide having at least 85% sequence identity with SEQ ID NO: 2 or which hybridizes under stringent conditions with SEQ ID NO: 1, the DNA sequence comprising at least one human beta casein intron sequence, and the system comprising a 5'-flanking sequence of a milk protein gene other than a casein gene, capable of mediating expression of said DNA sequence.

2. A method for producing a polypeptide having the amino acid sequence SEQ ID NO: 2 or an analogue or variant thereof which has the calcium binding activity of human beta casein, or opioid activity, or angiotensin converting enzyme (ACE) inhibitory activity, or a combination of any two or three of these activities, the method comprising introducing an expression system as claimed in claim 1 into the genome of a mouse in such a way that the DNA encoding the polypeptide is expressed in a mammary gland of the mouse, and collecting the polypeptide secreted from the mammary gland.

3. A method for producing a transgenic mouse capable of expressing a polypeptide having at least 85% sequence identity with SEQ ID NO: 2 or which hybridizes under stringent conditions with SEQ ID NO: 1, said method comprising chromosomally incorporating a DNA sequence comprising a polypeptide coding sequence encoding the polypeptide, at least one human beta casein intron sequence, and a promoter of a milk protein gene other than a casein gene, operatively associated with said coding sequence, into the genome of a mouse, whereby the polypeptide is expressed.

4. A transgenic mouse whose germ cells and somatic cells contain a DNA construct which comprises at least one human beta casein intron sequence and a promoter of a milk protein gene other than a casein gene, and a sequence which encodes a polypeptide having at least 85% sequence identity with SEQ ID NO: 2 or which hybridizes under stringent conditions with SEQ ID NO: 1, as a result of chromosomal incorporation of the construct into the mouse genome, or into the genome of an ancestor of said mouse, so that the DNA construct is expressed in a mammary gland of the transgenic mouse.

5. A transgenic mouse as claimed in claim 4 wherein the DNA construct is present within a mouse milk protein gene.

6. A method for supplementing a human infant formula comprising adding to said infant formula a polypeptide having the amino acid sequence of SEQ ID NO: 2 or an analogue or variant thereof which has the calcium binding activity of human beta casein, or opioid activity, or angiotensin converting enzyme (ACE) inhibitory activity, or a combination of any two or three of these activities, prepared by introducing an expression system according to claim 1 into the genome of a mouse in such a way that the DNA sequence which encodes a polypeptide having at least 85% sequence identity with SEQ ID NO: 2 or which hybridizes under stringent conditions with SEQ ID NO: 1, and comprises at least one human beta casein intron sequence, is capable of being expressed in a mammary gland of the mouse, obtaining expression of the polypeptide by said mouse, harvesting and optionally purifying the polypeptide expressed by said mouse.

7. An expression system according to claim 1 wherein the milk protein gene is a whey acidic protein gene.

Details for Patent 5,739,407

Applicant Tradename Biologic Ingredient Dosage Form BLA Number Approval Date Patent No. Assignee Estimated Patent Expiration Status Orphan Source
Schering INTRON A interferon alfa-2b VIAL 103132 001 1986-06-04 ⤷  Free Forever Trial Symbicom aktiebolag (Umea, SE) 2015-04-14 RX search
Schering INTRON A interferon alfa-2b VIAL 103132 002 1986-06-04 ⤷  Free Forever Trial Symbicom aktiebolag (Umea, SE) 2015-04-14 RX search
Schering INTRON A interferon alfa-2b VIAL 103132 003 1986-06-04 ⤷  Free Forever Trial Symbicom aktiebolag (Umea, SE) 2015-04-14 RX search
>Applicant >Tradename >Biologic Ingredient >Dosage Form >BLA >Number >Approval Date >Patent No. >Assignee >Estimated Patent Expiration >Status >Orphan >Source

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