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Claims for Patent: 6,660,477

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Summary for Patent: 6,660,477
Title: Method for the determination of data for the preparation of the diagnosis of phakomatosis
Abstract:The invention concerns a method for the determination of data for the preparation of presymptomatic or prenatal diagnosis of phakomatosis, in particular, a tumor suppressor gene disease, in a high-risk patient, in particular of neurofibromatosis, comprising the steps of: making available the tumor material from a person afflicted with the tumor suppressor gene disease, who is a relative of the high-risk patient; isolating tumor DNA from the tumor in the relative; isolating blood DNA from the blood of the relative; amplifying polymorphous DNA microsatellite markers from the tumor and the blood; separating the markers by length; observing the lengths of the markers; comparing the markers from the blood and the tumor; examining for a loss of alleles; optionally, comparing amplified markers from a second tumor of the relative; and amplifying polymorphous DNA microsatellite markers from the blood of an offspring and separating and observing the markers.
Inventor(s): Kluwe; Lan (Hamburg, DE)
Assignee: Von Recklinghausen Gesellschaft E.V. (Hamburg, DE)
Application Number:09/893,237
Patent Claims:1. A method for determining whether an offspring of an individual afflicted with neurofibromatosis has an increased risk of developing neurofibromatosis comprising the steps of: a. amplifying one or more polymorphous DNA microsatellite markers for neurofibromatosis from a tumor of the afflicted individual; b. amplifying the one or more polymorphous DNA microsatellite markers from the blood of the afflicted individual; c. comparing the amount and length of the one or more amplified polymorphous DNA microsatellite markers from steps (a) and (b); d. establishing the loss of an allele (Loss of Heterozygosity) in the tumor of the afflicted individual, based on the comparison in step (c); e. amplifying the one or more polymorphous DNA microsatellite markers from the blood of an offspring of the afflicted individual; and f. determining which allele of the afflicted individual was inherited by the offspring, wherein inheritance of the allele that is retained in the tumor of the afflicted individual indicates an increased risk of developing neurofibromatosis.

2. The method according to claim 1, wherein the one or more polymorphous DNA microsatellite markers amplified from the blood and the tumor of the afflicted individual are compared by length.

3. The method according to claim 1, wherein the offspring is not exhibiting symptoms of neurofibromatosis.

4. The method according to claim 1, wherein the offspring is a prenatal individual.

5. The method according to claim 1, further comprising the step of amplifying two or more different polymorphous DNA microsatellite markers.

6. A method for determining whether an offspring of an individual afflicted with neurofibromatosis has an increased risk of developing neurofibromatosis comprising the steps of: a) making available tumor material of the afflicted individual, b) making available blood of the afflicted individual, c) isolating the tumor DNA from the tumor material of the afflicted individual, d) isolating the blood DNA from the blood of the afflicted individual, e) amplifying one or more polymorphous DNA microsatellite markers for neurofibromatosis genes from the tumor material, f) amplifying the one or more polymorphous DNA microsatellite markers from the blood, g) separating by amount and length the polymorphous DNA microsatellite markers from the tumor material, h) separating by amount and length the polymorphous DNA microsatellite markers from the blood, i) observing the amount and length of the polymorphous DNA microsatellite markers from the tumor material, j) observing the amount and length of the polymorphous DNA microsatellite markers from the blood, k) determining an allele that is lost in the tumor material, l) amplifying the polymorphous DNA microsatellite markers from the blood of an offspring of the afflicted individual; and m) determining which allele of the afflicted individual was inherited by the offspring based on steps (k) and (l), wherein inheritance of the allele retained in the tumor indicates an increased risk of developing neurofibromatosis.

7. The method according to claim 1 or claim 6, wherein the polymorphous DNA microsatellite marker has a length of up to approximately 300 bp.

8. The method according to claim 5 or claim 6, wherein at least three different polymorphous DNA microsatellite markers are used.

9. The method according to claim 1 or claim 6, wherein the marker is a neurofibromatosis gene flanking marker or intragenic marker.

10. The method according to claim 9, wherein at least one of the markers is located in intron 27 of the neurofibromatosis type 1 gene.

11. The method according to claim 9, wherein at least one of the markers is located in intron 38 of the neurofibromatosis type 1 gene.

12. The method according to claim 9, wherein, the marker is selected from the group consisting of CRYB2, D22S275, NF2CA3, D22S268 and D22S430.

13. The method according to claim 1 or claim 6, further comprising amplifying the one or more markers in at least one additional tumor of the afflicted individual.

14. The method according to claim 1 or claim 6, further comprising the steps of amplifying the one or more polymorphous DNA microsatellite markers from the blood of an unaffected relative of the offspring and observing the amplified marker DNA.

15. The method according to claim 6, wherein at least two different polymorphous DNA microsatellite markers are used.

16. The method according to claim 5 or claim 6, wherein at least four different polymorphous DNA microsatellite markers are used.

Summary for Patent:   Start Trial

Foriegn Application Priority Data
Foreign Country Foreign Patent Number Foreign Patent Date
00113607Jun 27, 2000

Details for Patent 6,660,477

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   Start Trial Von Recklinghausen Gesellschaft E.V. (Hamburg, DE) 2020-06-27 RX search
Schering INTRON A interferon alfa-2b VIAL 103132 002 1986-06-04   Start Trial Von Recklinghausen Gesellschaft E.V. (Hamburg, DE) 2020-06-27 RX search
Schering INTRON A interferon alfa-2b VIAL 103132 003 1986-06-04   Start Trial Von Recklinghausen Gesellschaft E.V. (Hamburg, DE) 2020-06-27 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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