Claims for Patent: 6,340,595

Summary for Patent: 6,340,595

Title:	High throughput screening of gene function using adenoviral libraries for functional genomics applications
Abstract:	Novel means and methods for their use are provided to determine the function of the product(s) of one or more sample nucleic acids. The sample nucleic acids are synthetic oligonucleotides, DNA, or cDNA and encode polypeptides, antisense nucleic acids, or GSEs. The sample nucleic acids are expressed in a host by a vehicle to alter at least one phenotype of the host. The altered phenotype(s) is/are identified as a means to assign a biological function to the product(s) encoded by the sample nucleic acid(s).
Inventor(s):	Vogels; Ronald (Linschoten, NL), Bout; Abraham (Moerkapelle, NL), van Es; Helmuth (Hoofddorp, NL), Schouten; Govert (Leiden, NL)
Assignee:	Galapagos Genomics N.V. (Mechelen, BE)
Application Number:	09/358,036
Patent Claims:	1. A library of a multitude of unique expressible nucleic acids, said library including: a multiplicity of compartments, each of said compartments consisting essentially of one or more adenoviral vector comprising at least one unique nucleic acid of said library in an aqueous medium, wherein said adenoviral vector is capable of introducing said nucleic acid into a host cell, is capable of expressing the product of said nucleic acid in said host cell, and is deleted in a portion of the adenoviral genome necessary for replication thereof in said host cell. 2. The library of claim 1, wherein the function of the expression product of all of said unique expressible nucleic acids is unknown at the time said library is first made. 3. The library of claim 1, wherein none of said compartments contain any adenoviral vector capable of replication except in a packaging cell containing said deleted portion of adenoviral genome. 4. The library of claim 1, wherein said host cell is a eucaryotic cell. 5. The library of claim 1, wherein at least one compartment comprises at least two adenoviral vectors. 6. The library of claim 1, wherein each of said compartments consists essentially of one said adenoviral vector. 7. The library of claim 3, wherein each of said compartments contains from about 0.01.times.10.sup.10 to about 10.times.10.sup.10 pfu of said adenoviral vector per ml of aqueous medium. 8. The library of claim 7, wherein each of said compartments further contains the cellular debris from packaging cell lysate. 9. The library of claim 3, wherein said adenoviral vector is a minimal vector. 10. The library of claim 9, wherein said minimal vector comprises an adenovirus encapsidation signal or a functional part, derivative and/or analogue thereof, and at least one copy of at least a functional part or a derivative of an adenoviral ITR. 11. The library of claim 3, wherein said adenoviral vector comprises adenoviral genomic sequence deleted for sequence encoding the E1-region proteins. 12. The library of claim 10 wherein said minimal vector further comprises an adeno-associated virus terminal repeat or a functional part, derivative and/or analogue thereof. 13. The library of claim 11, wherein said adenoviral vector is further deleted for sequence encoding the E2A-region proteins, or the E2B region proteins or the complete E2 region proteins. 14. The library of claim 1, wherein said adenoviral vector further comprises adenovirus genomic sequence encoding adenoviral fiber proteins from at least two serotypes of adenovirus. 15. A process for producing a library consisting of a multitude of unique expressible nucleic acids arranged in a multiplicity of compartments, each said compartment consisting essentially of replication deficient adenoviral vector comprising at least one of said unique nucleic acids, comprising: transfecting (a) a packaging cell harboring a first portion of the adenoviral genome integrated into its genome, with an admixture of (b) a nucleic acid delivery vehicle containing said unique nucleic acid operably linked to a promoter and further containing a second portion of the adenoviral genome, said second portion comprising at least one adenoviral ITR, and (c) a helper nucleic acid consisting essentially of a third portion of the adenoviral genome; wherein the sequence of said first portion of adenoviral genome does not overlap with the sequences of either the second or third portions of adenoviral genome; and wherein the first, second and third portions of adenoviral genome are arranged such that all adenoviral proteins essential for replication and encapsidation are capable of expression in said packaging cells. 16. A process for producing a library according to claim 15, wherein said sequences of said second and third portions of adenoviral genome at least partially overlap allow for homologous recombination between said delivery vehicle nucleic acid and said helper nucleic acid. 17. A process for producing a library according to claim 16, wherein the sequences of adenoviral genome contained in said vehicle nucleic acid, and said helper nucleic acid, do no contain any portion of the E1 region. 18. A process for producing a library according to claim 17, wherein said packaging cell comprises a eucaryotic cell in which genome the E1 region of the adenoviral genome is integrated. 19. A process for producing a library of claim 18, wherein the sequences of said delivery vehicle and said helper nucleic acid are selected such that each is capable of being linearized by restriction enzymes that may be admixed therewith in the absence of further enzymatic restriction prior to transfection into said packaging cell. 20. The library according to claim 1, wherein said multiplicity of compartments comprises a multiwell format of at least 6 wells. 21. The library according to claim 20, wherein substantially each said well consists essentially of one or more said adenoviral vector comprising said unique nucleic acid that encodes a product of unknown function. 22. The library according to claim 1, wherein said library is configured to be made and used in a substantially automated process. 23. The library according to claim 20, wherein said multiplicity of compartments comprises a multiwell format of at least 96 wells. 24. The library according to claim 23, wherein each well contains cellular debris from eucaryotic packaging cell lysate. 25. The library of claim 24, wherein none of said wells contains adenoviral vector capable of replication except in a packaging cell containing said deleted portion of adenoviral genome. 26. The library of claim 25, wherein each of said wells contains from about 0.01.times.10.sup.10 to about 10.times.10.sup.10 pfu of said adenoviral vector per ml of aqueous medium. 27. The library of claim 1, wherein each of said unique nucleic acids is derived from a member of a population of nucleic acids, said population selected from the group consisting of naturally occurring populations of messenger RNA, DNAs, cDNAs, genes, ESTs, or genetic suppressor elements, and synthetic oligonucleotides, and antisense nucleic acids. 28. The library of claim 8, wherein the contents of each said compartment is capable of transfecting said host cell and expressing the product of each said unique nucleic acid in said host cell. 29. The library of claim 28, wherein each said compartment is capable of providing from about 400 to about 4000 aliquots of said adenoviral vector. 30. The library of claim 26, wherein the contents of each said well is capable of transfecting said host cell and expressing the product of each said unique nucleic acid in said host cell. 31. The library of claim 30, each said well is capable of providing from about 400 to about 4000 aliquots of said adenoviral vector. 32. The library of claim 10, wherein said minimal vector comprises an regulatable promoter operably linked to said unique nucleic acid. 33. The library of claim 11, wherein said adenoviral vector comprises an regulatable promoter operably linked to said unique nucleic acid. 34. The library of claim 11 or 13, wherein said adenoviral vector is further deleted for the adenoviral E3-region or a functional part thereof. 35. The library of claim 34, wherein said adenoviral vector is further deleted for the adenoviral E4-region or a functional part thereof. 36. The library of claim 32, wherein said promoter is repressed by an adenoviral E1 gene product. 37. The library of claim 33, wherein said promoter is repressed by an adenoviral E1 gene product. 38. The library of claim 36, wherein said promoter is an AP1 dependent promoter. 39. The library of claim 37, wherein said promoter is an AP1 dependent promoter. 40. A process for producing a library of claim 18, wherein said delivery vehicle comprises one or more restriction site sequence capable of cleavage by an enzyme that does not digest sequences coded for by said unique nucleic acids. 41. A process for producing a library of claim 40, wherein said restriction site sequence comprises a recognition sequence for a rare-cutting restriction endonuclease or an intron-encoded endonuclease. 42. A library according to claim 1, wherein said adenoviral vector is packaged into an adenoviral capsid. 43. A method according to claim 15, wherein said packaging cells are PER.C6 cells or derived from PER.C6 cells. 44. A method according to claim 43, wherein said cells include adenoviral genome sequence of the E2 region.

Details for Patent 6,340,595

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		2018-06-12
Merck Sharp & Dohme Corp.	INTRON A	interferon alfa-2b	For Injection	103132			2018-06-12
Merck Sharp & Dohme Corp.	INTRON A	interferon alfa-2b	Injection	103132			2018-06-12