Capsules Containing Transiently Transfected Cells, Method for Preparing Same and Uses Thereof
The invention concerns a capsule containing cells that are transiently transfected with a gene of interest and entrapped within a biocompatible polymer membrane, a method of preparing those capsules, a method for in vivo assessing an activity of the protein secreted by the gene of interest, which comprises administering to a multicellular organism the capsule and detecting an activity, a pharmaceutical composition comprising such a capsule, and the use thereof for the administration of a protein of interest to a subject.
1 - 26 . (canceled)
27 . A composition of matter comprising a capsule containing cells that are transiently transfected with a gene of interest and entrapped within a biocompatible polymer membrane.
28 . The composition of matter according to claim 27 , wherein the cells are animal cells.
29 . The composition of matter according to claim 28 , wherein said animal cells are mammalian.
30 . The composition of matter according to claim 27 , wherein the gene of interest is fused to a signal sequence for secretion of the protein, inserted into an expression cassette or inserted into a plasmid.
31 . The composition of matter according to claim 27 , wherein the biocompatible polymer membrane is composed of alginate-poly-L-lysine-alginate (APA).
32 . The composition of matter according to claim 27 , wherein the biocompatible polymer membrane has pores with a cut-off size of 90 kDa to 30 kDa or 80 kDa to 60 kDa.
33 . The composition of matter according to claim 31 , wherein the biocompatible polymer membrane has pores with a cut-off size of 90 kDa to 30 kDa or 80 kDa to 60 kDa.
34 . The composition of matter according to claim 27 , wherein the capsule has a mean diameter of 100 to 1500 μm, 250 to 600 μm or 440 to 530 μm.
35 . The composition of matter according to claim 27 , wherein the capsule has been maintained under low-shear microgravity conditions.
36 . The composition of matter according to claim 27 , further comprising one or more pharmaceutically acceptable carriers, diluents or excipients.
37 . The composition of matter according to claim 27 , wherein the gene of interest is coding for an antigen/immunogen and/or an adjuvant.
38 . The composition of matter according to claim 37 , wherein the antigen/immunogen is a bacterial, viral, fungal, parasitic or tumor antigen.
39 . The composition of matter according to claim 27 , wherein the capsule comprises at least two types of cells, each transfected with a gene encoding one or more antigen and/or one or more adjuvant and wherein the two genes are not identical.
40 . A method of preparing a capsule comprising the steps of transiently transfecting cells with a gene of interest and encapsulating the transiently transfected cells.
41 . The method according to claim 40 , further comprising maintaining the capsule under low-shear microgravity gravity conditions.
42 . A method for assessing an in vivo activity of the protein expressed and secreted by a gene of interest, which comprises administering a capsule according to claim 27 to a multicellular organism and detecting an activity of said protein.
43 . The method according to claim 42 , wherein the multicellular organism is a mammal.
44 . The method according to claim 43 , wherein the mammal is selected from the group consisting of mouse, rats, dogs, goats, sheep, cows and monkeys.
45 . The method according to claim 43 , wherein administration of the capsule is performed by i.p. injection of a mammal.
46 . The method according to claim 43 , wherein the in vivo activity to be detected is completely induced within a period of 3 to 14 days, or within a period of 3 to 5 days after administration of said protein to said mammal.
47 . The method according to claim 46 , wherein said mammal is a mouse with Concanavalin A (ConA) induced liver toxicity.
48 . A method of administering a protein of interest to a subject comprising administering a composition of matter comprising a capsule according to claim 27 that expresses a protein of interest to a subject.
49 . The method according to claim 48 , wherein the protein of interest is an antigen for immunization or vaccination.
50 . The method according to claim 48 , wherein the subject is selected from the group consisting of humans; animals kept for research purposes; livestock; and companion animals.
51 . A kit comprising a composition of matter according to claim 27 and means for the application of said composition of matter to a subject.