Abstract: The current teachings are directed to novel virus free cells lines derived from virus-contaminated staring material, such as an organism or a cell line. Methods for obtaining virus free cell lines obtained from virus-contaminated starting material are also provided. Exemplary virus free cell lines include: novel cell lines derived from a Spodoptera frugiperda cell line contaminated with Sf-rhabdovirus, wherein the novel cell lines lack Sf-rhabdovirus; and novel cell lines derived from a Trichoplusia ni cell line contaminated with an alphanodavirus, wherein the novel cell line lacks an alphanodavirus.

Abstract: The current teachings are directed to virus free cells lines derived from virus-contaminated starting material, such as an organism or a cell line. Methods for obtaining virus free cell lines obtained from virus-contaminated starting material are also provided. Exemplary virus free cell lines include: cell lines derived from a Spodoptera frugiperda cell line contaminated with Sf-rhabdovirus, wherein the cell lines lack Sf-rhabdovirus; and cell lines derived from a Trichoplusia ni cell line contaminated with an alphanodavirus, wherein the cell line lacks an alphanodavirus.

Abstract: The current teachings are directed to novel virus free cells lines derived from virus-contaminated staring material, such as an organism or a cell line. Methods for obtaining virus free cell lines obtained from virus-contaminated starting material are also provided. Exemplary virus free cell lines include: novel cell lines derived from a Spodoptera frugiperda cell line contaminated with Sf-rhabdovirus, wherein the novel cell lines lack Sf-rhabdovirus; and novel cell lines derived from a Trichoplusia ni cell line contaminated with an alphanodavirus, wherein the novel cell line lacks an alphanodavirus.

Abstract: The present invention relates to methods of facilitating the expression of recombinant polypeptides from cells, extracellular fluids, extracellular fibers, or any combination thereof, obtained from transgenic insect cells and larvae comprising a bacterial GlcNAc-6-P 2?-epimerase (GNPE), which is capable of converting N-acetyl-D-glucosamine-6-phosphate (GlcNAc-6-P) to N-acetyl-D-mannosamine-6-phosphate (ManNAc-6-P). The invention relates to methods to promote efficient glycoconjugate sialylation, by providing simpler ways to produce large intracellular pools of sialic acid precursors. The invention is also directed to nucleic acids, vectors, and cells comprising nucleic acids encoding polypeptides involved in the synthesis of sialic acid precursors, and cells in combination with nucleic acids encoding glycosyltransferases, including sialyltransferases, to facilitate the production of humanized recombinant glycoproteins.

Abstract: The present invention relates to new methods to promote sialylation of glycoconjugates, including recombinant glycoproteins, in glycoconjugate production systems. The invention relates to methods to promote efficient glycoconjugate sialylation in recombinant expression systems, by providing simpler and more economical ways to produce large intracellular pools of sialic acid precursors. The invention is directed to nucleic acids, vectors, and cells harboring vectors comprising nucleic acids encoding enzymes involved in the synthesis of sialic acid precursors, and cells harboring these nucleic acids in combination with nucleic acids encoding glycosyltransferases, including sialyltransferases, to facilitate the production of humanized recombinant glycoproteins in bacterial, fungal, plant, and animal cell expression systems.

Abstract: The present invention relates to methods of facilitating the expression of recombinant polypeptides from cells, extracellular fluids, extracellular fibers, or any combination thereof, obtained from transgenic insect cells and larvae comprising a bacterial GlcNAc-6-P 2?-epimerase (GNPE), which is capable of converting N-acetyl-D-glucosamine-6-phosphate (GlcNAc-6-P) to N-acetyl-D-mannosamine-6-phosphate (ManNAc-6-P). The invention relates to methods to promote efficient glycoconjugate sialylation, by providing simpler ways to produce large intracellular pools of sialic acid precursors. The invention is also directed to nucleic acids, vectors, and cells comprising nucleic acids encoding polypeptides involved in the synthesis of sialic acid precursors, and cells in combination with nucleic acids encoding glycosyltransferases, including sialyltransferases, to facilitate the production of humanized recombinant glycoproteins.

Abstract: Methods for manipulating carbohydrate processing pathways in cells of interest are provided. Methods are directed at manipulating multiple pathways involved with the sialylation reaction by using recombinant DNA technology and substrate feeding approaches to enable the production of sialylated glycoproteins in cells of interest. These carbohydrate engineering efforts encompass the implementation of new carbohydrate bioassays, the examination of a selection of insect cell lines and the use of bioinformatics to identify gene sequences for critical processing enzymes. The compositions comprise cells of interest producing sialylated glycoproteins. The methods and compositions are useful for heterologous expression of glycoproteins.

Abstract: Methods for manipulating carbohydrate processing pathways in cells of interest are provided. Methods are directed at manipulating multiple pathways involved with the sialylation reaction by using recombinant DNA technology and substrate feeding approaches to enable the production of sialylated glycoproteins in cells of interest. These carbohydrate engineering efforts encompass the implementation of new carbohydrate bioassays, the examination of a selection of insect cell lines and the use of bioinformatics to identify gene sequences for critical processing enzymes. The compositions comprise cells of interest producing sialylated glycoproteins. The methods and compositions are useful for heterologous expression of glycoproteins.

Abstract: The present invention relates to new methods to promote sialylation of glycoconjugates, including recombinant glycoproteins, in glycoconjugate production systems. The invention relates to methods to promote efficient glycoconjugate sialylation in recombinant expression systems, by providing simpler and more economical ways to produce large intracellular pools of sialic acid precursors. The invention is directed to nucleic acids, vectors, and cells harboring vectors comprising nucleic acids encoding enzymes involved in the synthesis of sialic acid precursors, and cells harboring these nucleic acids in combination with nucleic acids encoding glycosyltransferases, including sialyltransferases, to facilitate the production of humanized recombinant glycoproteins in bacterial, fungal, plant, and animal cell expression systems.

Abstract: Methods for manipulating carbohydrate processing pathways in cells of interest are provided. Methods are directed at manipulating multiple pathways involved with the sialylation reaction by using recombinant DNA technology and substrate feeding approaches to enable the production of sialylated glycoproteins in cells of interest. These carbohydrate engineering efforts encompass the implementation of new carbohydrate bioassays, the examination of a selection of insect cell lines and the use of bioinformatics to identify gene sequences for critical processing enzymes. The compositions comprise cells of interest producing sialylated glycoproteins. The methods and compositions are useful for heterologous expression of glycoproteins.

Abstract: This invention relates, e.g., to transgenic insects, or progeny thereof, whose cells contain at least one genomically integrated, expressible, nucleic acid encoding two or more of a set of Nglycosylation enzymes that can glycosylate a heterologous protein with a mammalianized (e.g., humanized) glycosylation pattern. The glycosylation genes are preferably expressed in the insect cells in catalytic amounts. Also described are methods to use such a transgenic insect to produce heterologous, mammalianized polypeptides of interest.

Abstract: The invention includes a nucleic acid having a sequence at least 98% homologous to SEQ.ID.NO: 1, which encodes the a subunit of canine TSH. The invention also includes a nucleic acid having a sequence at least 98% homologous to SEQ.ID.NO: 2, which encodes the ? subunit of canine TSH. The invention also includes a method of producing an rcTSH subunit by expressing a nucleic acid having a sequence of SEQ.ID.NO: 1 and a nucleic acid having a sequence of SEQ.ID.NO: 2 in a transgenic insect cell modified to sialylate proteins and producing a sialylated rcTSH subunit. The insect cell may be a lepidopteran cell. The rcTSH may be used for diagnosis and treatment. It may be used to diagnose canine hypothyroidism.