Abstract: This disclosure relates to inexpensive and efficient methods of preparing egg white (e.g., obtained from eggs laid by transgenic chickens) for bulk chromatographic isolation of proteins (e.g., recombinant proteins) from the egg white, as well as method of filtering acidified egg white and methods of isolating proteins from the egg white.

Abstract: The present invention is related a method of purifying a heterologous protein from an egg white. In some embodiments, the disclosure is directed to a method of purifying a heterologous protein from an egg white comprising the heterologous protein, the method comprising, (a) adjusting the pH of the egg white to a pH of 5.8 to 6.5 to form a pH-adjusted egg white; (b) filtering the pH-adjusted egg white of (a) and collecting a first filtrate; (c) subjecting the first filtrate of (b) to a hydrophobic interaction chromatography matrix, and collecting a first eluate comprising the heterologous protein; (d) adjusting the pH of the first eluate of (c) to a pH of 5.0 to 5.6 to form a pH-adjusted eluate; (e) filtering the pH-adjusted eluate to obtain a second filtrate; (f) adjusting the pH of the second filtrate to a pH of 3.0 to 4.0 to form a pH-adjusted second filtrate; (g) neutralizing the pH-adjusted second filtrate of (f) to a pH of 5.0 to 8.

Abstract: Methods of producing an exogenous protein in a bird egg are provided according to aspects of the present invention which include: providing transfected avian primordial germ cells by forming a complex of Cas protein and a guide nucleotide sequence in the avian primordial germ cells, whereby a donor DNA sequence is inserted into genomic DNA of the avian primordial germ cells under transcriptional control of a regulatory element that directs tissue-specific expression of the exogenous protein; introducing the transfected avian primordial germ cells into a population of recipient bird embryos and incubating the recipient bird embryos, generating germline chimera birds; obtaining a heterozygote and/or homozygote transgenic bird by breeding the germline chimera bird; and isolating the exogenous protein from an egg laid by a female transgenic bird.

Abstract: The present invention is directed to isolated recombinant human ADA2 proteins, ADA2 biologically active fragments, and ADA2 fusion proteins. The proteins of the invention can be surprisingly used to restore ADA2 activity in subjects having loss-of-function mutations in ADA2 and increase differentiation of monocytes into macrophages, e.g., M2 macrophages, e.g., M2c macrophages, stimulate CD4+ T cell proliferation, and increase endothelial cell development. More specifically, the ADA2 proteins, ADA2 biologically active fragments, and ADA2 fusion proteins of the invention can be used to treat subjects having ADA2-associated diseases or disorders, including but not limited to, polyarteritis nodosa, Sneddon Syndrome, vasculitis, ischemic stroke, hemorrhagic stroke, lacunar stroke, aneurysm in the celiac artery, skin rash, skin necrosis, livedo racemosa, hepatosplenomegaly, organ failure, retinal artery occlusion, optic nerve atrophy, diplopia, cranial nerve palsy, strabismus, and low serum IgM.

Abstract: The present invention is related a method of purifying a heterologous protein from an egg white. In some embodiments, the disclosure is directed to a method of purifying a heterologous protein from an egg white comprising the heterologous protein, the method comprising, (a) adjusting the pH of the egg white to a pH of 5.8 to 6.5 to form a pH-adjusted egg white; (b) filtering the pH-adjusted egg white of (a) and collecting a first filtrate; (c) subjecting the first filtrate of (b) to a hydrophobic interaction chromatography matrix, and collecting a first eluate comprising the heterologous protein; (d) adjusting the pH of the first eluate of (c) to a pH of 5.0 to 5.6 to form a pH-adjusted eluate; (e) filtering the pH-adjusted eluate to obtain a second filtrate; (f) adjusting the pH of the second filtrate to a pH of 3.0 to 4.0 to form a pH-adjusted second filtrate; (g) neutralizing the pH-adjusted second filtrate of (f) to a pH of 5.0 to 8.

Abstract: This disclosure relates to inexpensive and efficient methods of preparing egg white (e.g., obtained from eggs laid by transgenic chickens) for bulk chromatographic isolation of proteins (e.g., recombinant proteins) from the egg white, as well as method of filtering acidified egg white and methods of isolating proteins from the egg white.

Abstract: The present invention is directed to isolated recombinant human ADA2 proteins, ADA2 biologically active fragments, and ADA2 fusion proteins. The proteins of the invention can be surprisingly used to restore ADA2 activity in subjects having loss-of-function mutations in ADA2 and increase differentiation of monocytes into macrophages, e.g., M2 macrophages, e.g., M2c macrophages, stimulate CD4+ T cell proliferation, and increase endothelial cell development. More specifically, the ADA2 proteins, ADA2 biologically active fragments, and ADA2 fusion proteins of the invention can be used to treat subjects having ADA2-associated diseases or disorders, including but not limited to, polyarteritis nodosa, Sneddon Syndrome, vasculitis, ischemic stroke, hemorrhagic stroke, lacunar stroke, aneurysm in the celiac artery, skin rash, skin necrosis, livedo racemosa, hepatosplenomegaly, organ failure, retinal artery occlusion, optic nerve atrophy, diplopia, cranial nerve palsy, strabismus, and low serum IgM.

Abstract: This disclosure relates to inexpensive and efficient methods of preparing egg white (e.g., obtained from eggs laid by transgenic chickens) for bulk chromatographic isolation of proteins (e.g., recombinant proteins) from the egg white, as well as methods of filtering acidified egg white and methods of isolating proteins from the egg white.

Abstract: The present invention provides compositions comprising an isolated mixture of recombinant human NaGlu proteins in which a substantial amount of the NaGlu proteins in the mixture has increased levels of phosphorylated mannose that confer the proteins to be efficiently internalized into human cells. The present invention also provides methods of producing such mixture of NaGlu proteins, vectors used in transgenesis and expression, host cells harboring such vectors, and methods of isolating and purifying the mixture of NaGlu proteins. The invention further provides methods of treating NaGlu associated diseases.

Abstract: The present invention provides compositions comprising an isolated mixture of recombinant human NaGlu proteins in which a substantial amount of the NaGlu proteins in the mixture has increased levels of phosphorylated mannose that confer the proteins to be efficiently internalized into human cells. The present invention also provides methods of producing such mixture of NaGlu proteins, vectors used in transgenesis and expression, host cells harboring such vectors, and methods of isolating and purifying the mixture of NaGlu proteins. The invention further provides methods of treating NaGlu associated diseases.

Abstract: This disclosure relates to inexpensive and efficient methods of preparing egg white (e.g., obtained from eggs laid by transgenic chickens) for bulk chromatographic isolation of proteins (e.g., recombinant proteins) from the egg white, as well as methods of filtering acidified egg white and methods of isolating proteins from the egg white.

Abstract: The present invention provides compositions comprising an isolated mixture of recombinant human NaGlu proteins in which a substantial amount of the NaGlu proteins in the mixture has increased levels of phosphorylated mannose that confer the proteins to be efficiently internalized into human cells. The present invention also provides methods of producing such mixture of NaGlu proteins, vectors used in transgenesis and expression, host cells harboring such vectors, and methods of isolating and purifying the mixture of NaGlu proteins. The invention further provides methods of treating NaGlu associated diseases.

Abstract: The invention relates to transgenic avians which produce antibodies in the egg white by introducing a nucleic acid sequence into the genome of an avian embryo wherein the nucleic acid sequence comprises a nucleotide sequence encoding an antibody and to the antibodies and to methods related thereto.

Abstract: Methods of producing protein using a recombinant ovomucoid gene expression controlling region operably linked to one or more useful amino acid coding sequences.

Abstract: The present invention provides for methods of producing transgenic avians which may include delivering a heterologous nucleic acid to oviduct tissue of an avian wherein the nucleic acid enters a cell of the oviduct tissue and is expressed.

Abstract: The invention includes methods of making transgenic avians by introducing a nucleic acid sequence into the genome of an avian embryo wherein the nucleic acid sequence comprises a nucleotide sequence encoding a heterologous protein, developing the avian embryo to hatch, developing the hatched chick to sexual maturity and obtaining offspring from the sexually mature hatched chick which produce egg white containing the heterologous protein.

Abstract: The invention provides methods of purifying proteins, e.g., cytokines, from egg whites.

Abstract: The present invention provides isolated and recombinant avian nucleic acid molecules comprising at least one avian MAR and an avian nucleic acid sequence encoding an ovalbumin transcriptional regulatory region. The isolated nucleic acid of the present invention is useful for reducing chromosomal positional effects upon the transcription of a transgene operably linked to the ovalbumin transcriptional regulatory region and transfected into a recipient avian cell The recombinant nucleic acid molecules of the present invention may further comprise a polyadenylation signal sequence or an avian 3? domain, and optionally, an internal ribosome entry site for expression of an operably linked heterologous nucleic acid insert in a transfected avian cell.

Abstract: The invention includes delivering to an avian a nucleic acid molecule which includes nucleotide sequence of an ovalbumin gene expression controlling region and a heterologous coding sequence operably linked to the gene expression controlling region wherein the heterologous coding sequence is expressed in a cell of the avian.

Abstract: The present invention provides for methods of producing transgenic avians which may include delivering a heterologous nucleic acid to oviduct tissue of an avian wherein the nucleic acid enters a cell of the oviduct tissue and is expressed.