Abstract: Methods for altering the N-glycosylation pattern of proteins in higher plants are provided. The methods comprise introducing into the plant a recombinant RNAi construct that provides for the inhibition of expression of ?1,3-fucosyltransferase (FucT) and ?1,2-xylosyltransferase (XylT) in a plant. Use of these RNAi constructs to inhibit or suppress expression of both of these enzymes, and isoforms thereof, advantageously provides for the production of endogenous and heterologous proteins having a “humanized” N-glycosylation pattern without impacting plant growth and development. Stably transformed higher plants having this protein N-glycosylation pattern are provided. Glycoprotein compositions, including monoclonal antibody compositions, having substantially homogeneous glycosylation profiles, and which are substantially homogeneous for the G0 glycoform, are also provided.

Abstract: The present invention encompasses influenza vaccines, in particular avian influenza vaccines. The vaccine may be a subunit vaccine based on the hemagglutinin of influenza. The hemagglutinin may be expressed in plants including duckweed. The invention also encompasses recombinant vectors encoding and expressing influenza antigens, epitopes or immunogens which can be used to protect animals against influenza. It encompasses also a vaccination regimen compatible with the DIVA strategy, including a prime-boost scheme using vector and subunit vaccines.

Abstract: The invention pertains to anti-CD30 antibodies that lack fucosyl and xylosyl residues. The antibodies of the invention exhibit increased antibody-dependent cellular cytotoxicity (ADCC) activity, including the ability to lyse CD30-expressing cell lines that are not lysed by the fucosylated and xylosylated form of the antibodies. The invention also provides host cells that express the anti-CD30 antibodies that lack fucosyl and xylosyl residues, wherein the host cells are deficient for a fucosyltransferase and a xylosyltransferase. Methods of using the antibodies to inhibit the grown of CD30+ cells, such as tumor cells, are also provided.

Abstract: Compositions and methods for producing aglycosylated plasminogen (PLG) polypeptides and fragments and variants thereof are provided. Compositions of the invention include isolated nucleic acid molecules encoding aglycosylated PLG polypeptides in which the asparagine (Asn) residue corresponding to residue Asn-289 of the mature human PLG polypeptide has been substituted with an amino acid residue that does not support N-linked glycosylation at that position of the PLG polypeptide, as well as the aglycosylated PLG polypeptides encoded thereby. Expression constructs comprising these PLG-encoding nucleic acid molecules and transgenic plants comprising these expression constructs are also provided. Methods of the invention comprise introducing a PLG-encoding nucleic acid molecule of the invention into a plant of interest and culturing the plant under conditions to produce the aglycosylated PLG polypeptide.

Abstract: Compositions and methods for regulating expression of nucleotide sequences of interest in a plant are provided. Compositions include novel nucleic acid molecules, and variants and fragments thereof, for expression control elements isolated from the Lemnaceae ubiquitin, r-histone and chitinase genes. A method for expressing a nucleotide sequence of interest in a plant using the expression control elements disclosed herein is further provided. The method includes introducing into a plant or plant cell or nodule an expression construct comprising an expression control element of the present invention operably linked to a nucleotide sequence of interest. In particular, the compositions and methods find use in enhancing expression of nucleotide sequences of interest in duckweed. Also provided is a novel Lemnaceae signal peptide-encoding sequence and the signal peptide encoded thereby.

Abstract: The present invention provides methods and compositions for the production of recombinant plasminogen, microplasminogen, and fragments thereof in a duckweed expression system. It is the novel finding of the present invention that a duckweed expression system may be used to produce high levels of plasminogen and microplasminogen. The duckweed-produced plasminogen and microplasminogen can be activated to produce a polypeptide having protease activity. Thus, the invention encompasses methods for the expression of plasminogen, microplasminogen, and fragments thereof in duckweed, duckweed plants that are transformed with expression cassettes for the expression of plasminogen, microplasminogen, and fragments thereof, and nucleic acids comprising nucleotide sequences encoding plasminogen, microplasminogen, and fragments thereof, where these nucleotide sequences are modified to enhance their expression in duckweed.

Abstract: Methods for altering the N-glycosylation pattern of proteins in higher plants are provided. The methods comprise introducing into the plant a recombinant construct that provides for the inhibition of expression of ?1,3-fucosyltransferase (FucT) and ?1,2-xylosyltransferase (XylT) in a plant. Use of these constructs to inhibit or suppress expression of both of these enzymes, and isoforms thereof, advantageously provides for the production of endogenous and heterologous proteins having a “humanized” N-glycosylation pattern without impacting plant growth and development. Stably transformed higher plants having this protein N-glycosylation pattern are provided. Glycoprotein compositions, including monoclonal antibody compositions, having substantially homogeneous glycosylation profiles, and which are substantially homogeneous for the G0 glycoform, are also provided.

Abstract: Methods for altering the N-glycosylation pattern of proteins in higher plants are provided. In some embodiments, the methods comprise introducing into a duckweed plant a recombinant RNAi construct that provides for the inhibition of expression of ?1,3-fucosyltransferase (FucT) and ?1,2-xylosyltransferase (XylT). Use of these RNAi constructs to inhibit or suppress expression of both of these enzymes, and isoforms thereof, advantageously provides for the production of endogenous and heterologous proteins having a “humanized” N-glycosylation pattern without impacting plant growth and development. Stably transformed higher plants, including duckweed plants, having this protein N-glycosylation pattern are provided. Glycoprotein compositions, including monoclonal antibody compositions, having substantially homogeneous glycosylation profiles, and which are substantially homogeneous for the G0 glycoform, are also provided.

Abstract: A bioreactor system according to one embodiment of the present invention is provided for producing and processing a biological material in an aseptic environment. For example, the system includes at least one culture tube configured to hold a liquid media containing a biological material and at least one flexible bag for promoting growth of the biological material therein. The liquid media and biological material are configured to be transferred from the culture tube to the flexible bag aseptically in an unclassified area. The system also includes at least one flexible harvest bag configured to be in flow communication with the flexible bag, wherein the flexible harvest bag is configured to separate biological material grown in the flexible bag from the liquid media aseptically in an unclassified area.

Abstract: The present invention encompasses influenza vaccines, in particular avian influenza vaccines. The vaccine may be a subunit vaccine based on the hemagglutinin of influenza. The hemagglutinin may be expressed in plants including duckweed. The invention also encompasses recombinant vectors encoding and expressing influenza antigens, epitopes or immunogens which can be used to protect animals against influenza. It encompasses also a vaccination regimen compatible with the DIVA strategy, including a prime-boost scheme using vector and subunit vaccines.

Abstract: The present invention provides methods and compositions for the production of recombinant plasminogen, microplasminogen, and fragments thereof in a duckweed expression system. It is the novel finding of the present invention that a duckweed expression system may be used to produce high levels of plasminogen and microplasminogen. The duckweed-produced plasminogen and microplasminogen can be activated to produce a polypeptide having protease activity. Thus, the invention encompasses methods for the expression of plasminogen, microplasminogen, and fragments thereof in duckweed, duckweed plants that are transformed with expression cassettes for the expression of plasminogen, microplasminogen, and fragments thereof, and nucleic acids comprising nucleotide sequences encoding plasminogen, microplasminogen, and fragments thereof, where these nucleotide sequences are modified to enhance their expression in duckweed.

Abstract: Compositions and methods for regulating expression of nucleotide sequences of interest in a plant are provided. Compositions include novel nucleic acid molecules, and variants and fragments thereof, for expression control elements isolated from the Lemnaceae ubiquitin, r-histone and chitinase genes. A method for expressing a nucleotide sequence of interest in a plant using the expression control elements disclosed herein is further provided. The method includes introducing into a plant or plant cell or nodule an expression construct comprising an expression control element of the present invention operably linked to a nucleotide sequence of interest. In particular, the compositions and methods find use in enhancing expression of nucleotide sequences of interest in duckweed. Also provided is a novel Lemnaceae signal peptide-encoding sequence and the signal peptide encoded thereby.

Abstract: The present invention provides methods and compositions for the production of recombinant plasminogen, microplasminogen, and fragments thereof in a duckweed expression system. It is the novel finding of the present invention that a duckweed expression system may be used to produce high levels of plasminogen and microplasminogen. The duckweed-produced plasminogen and microplasminogen can be activated to produce a polypeptide having protease activity. Thus, the invention encompasses methods for the expression of plasminogen, microplasminogen, and fragments thereof in duckweed, duckweed plants that are transformed with expression cassettes for the expression of plasminogen, microplasminogen, and fragments thereof, and nucleic acids comprising nucleotide sequences encoding plasminogen, microplasminogen, and fragments thereof, where these nucleotide sequences are modified to enhance their expression in duckweed.

Abstract: Methods, nucleic acid sequences, and transformed duckweed plant or duckweed nodule cultures for the expression and the secretion of biologically active polypeptides from genetically engineered duckweed are provided. Expression of recombinant polypeptides in duckweed is improved by modifying the nucleotide sequence of the expression cassette encoding the polypeptide for improved expression in duckweed. Recovery of biologically active polypeptides from duckweed is improved by linking the biologically active polypeptide to a signal peptide that directs the secretion of the polypeptide into the culture medium. Methods of producing recombinant monoclonal antibodies having effector function, and antibodies produced by these methods are provided.

Abstract: Compositions and methods for regulating expression of nucleotide sequences of interest in a plant are provided. Compositions include novel nucleic acid molecules, and variants and fragments thereof, for expression control elements isolated from the Lemnaceae ubiquitin, r-histone and chitinase genes. A method for expressing a nucleotide sequence of interest in a plant using the expression control elements disclosed herein is further provided. The method includes introducing into a plant or plant cell or nodule an expression construct comprising an expression control element of the present invention operably linked to a nucleotide sequence of interest. In particular, the compositions and methods find use in enhancing expression of nucleotide sequences of interest in duckweed. Also provided is a novel Lemnaceae signal peptide-encoding sequence and the signal peptide encoded thereby.

Abstract: The invention pertains to anti-CD30 antibodies that lack fucosyl and xylosyl residues. The antibodies of the invention exhibit increased antibody-dependent cellular cytotoxicity (ADCC) activity, including the ability to lyse CD30-expressing cell lines that are not lysed by the fucosylated and xylosylated form of the antibodies. The invention also provides host cells that express the anti-CD30 antibodies that lack fucosyl and xylosyl residues, wherein the host cells are deficient for a fucosyltransferase and a xylosyltransferase. Methods of using the antibodies to inhibit the growth of CD30 cells, such as tumor cells, are also provided.

Abstract: Glycan-optimized monoclonal antibodies that specifically bind CD20 antigen and which have improved effector function are provided. The anti-CD20 antibodies of the invention have a glycosylation pattern that results in an antibody composition having predominately the G0 glycoform, and thus comprise N-glycans that lack fucose (i.e., afucosylated) and galactose residues attached thereto. In some embodiments, these anti-CD20 antibodies comprise the light chain and heavy chain sequences of the rituximab anti-CD20 antibody, and thus represent afucosylated rituximab. Methods for producing these glycan-optimized anti-CD20 antibodies are also provided.

Abstract: Methods, nucleic acid sequences, and transformed duckweed plant or duckweed nodule cultures for the expression and the secretion of biologically active polypeptides from genetically engineered duckweed are provided. Expression of recombinant polypeptides in duckweed is improved by modifying the nucleotide sequence of the expression cassette encoding the polypeptide for improved expression in duckweed. Recovery of biologically active polypeptides from duckweed is improved by linking the biologically active polypeptide to a signal peptide that directs the secretion of the polypeptide into the culture medium. Methods of producing recombinant monoclonal antibodies having effector function, and antibodies produced by these methods are provided.