Large Scale Biology Corporation

Biosource Technologies, Inc.

Biosource Genetics Corporation

Other

Kentucky Bioprocessing, Inc.

Stephen J Garger

Thomas H Turpen

Michael J McCulloch

Laurence K Grill

R Barry Holtz

William O Dawson

Ann Myers Turpen

Jon Donson

Genadie G Sverlow

Michelle L Samonek-Potter

Terri I Cameron

Richard Barry Holtz

George L Granthan

George L Grantham

Monto H Kumagai

John A Lindbo

Gregory P Pogue

Guy R Della-Cioppa

Guy Richard della-Cioppa

Jonathan E Lawrence

Cynthia S Gross

Barry Bratcher

Miles R Chedekal

Jonathan Donson

Richard K Teague

Harriet Flannery Phillips

Miles R Chedekel

The present invention relates to foreign peptide sequences fused to recombinant plant viral structural proteins and a method of their production. Fusion proteins are economically synthesized in plants at high levels by biologically contained tobamoviruses. The fusion proteins of the invention have are useful as antigens for inducing the production of antibodies having desired binding properties, e.g., protective antibodies, or for use as vaccine antigens for the induction of protective immunity against the parvovirus. Feline parvovirus epitopes were fused to the N-terminus of the TMV coat protein, expressed inplants, extracted, purified, characterized and administered to animals, resulting in protective immunity.

Production of a parvovirus vaccine in plants as viral coat protein fusions

The present invention relates to a recombinant viral nucleic acid selected from a (+) sense, single stranded RNA virus possessing a native subgenomic promoter encoding for a first viral subgenomic promoter, a nucleic acid sequence that codes for a viral coat protein whose transcription is regulated by the first viral subgenomic promoter, a second viral subgenomic promoter and a second nucleic acid sequence whose transcription is regulated by the second viral subgenomic promoter. The first and second viral subgenomic promoters of the recombinant viral nucleic acid do not have homologous sequences relative to each other. The recombinant viral nucleic acid provides the particular advantage that it systemically transcribes the second nucleic acid in the host. Host organisms encompassed by the present invention include procaryotes and eucaryotes, particularly animals and plants. The present invention also relates to viruses containing the viral vectors which are infective, production cells which are capable of producing the viruses or parts thereof, a host infected by the viruses of the invention, the gene products produced by expression of the viral nucleic acids and a process for the production of a desired product by growing the infected hosts.

Recombinant viral nucleic acids

A flexible automated apparatus for isolating and purifying viruses, proteins and peptides of interest from a plant material is disclosed, the apparatus being applicable for large scale purification and isolation of such substances from plant material. The flexible automated apparatus provides an efficient apparatus for isolating viruses, proteins and peptides of interest with little waste material. The automated apparatus for isolating viruses, proteins and peptides of interest includes a grinding apparatus for homogenizing a plant to produce a green juice, a means for adjusting the pH of and heating the green juice, a means for separating the target species, either virus or protein/peptide, from other components of the green juice by one or more cycles of centrifugation, resuspension, and ultrafiltration, and finally purifying virus particles by such procedure as PEG-precipitation or purifying proteins and peptides by such procedures as chromatography and/or salt precipitation.

Flexible processing apparatus for isolating and purifying viruses, soluble proteins and peptides from plant sources

A method for extracting proteins from the intercellular space of plants is provided. The method is applicable to the large scale isolation of many active proteins of interest synthesized by plant cells. The method may be used commercially to recover recombinantly produced proteins from plant hosts thereby making the large scale use of plants as sources for recombinant protein production feasible.

Method for recovering proteins from the interstitial fluid of plant tissues

The invention relates to &#x3b1;-galactosidase truncated at the carboxy terminus and the production of enzymatically active recombinant human and animal lysosomal enzymes involving construction and expression of recombinant expression constructs comprising coding sequences of human or animal lysosomal enzymes in a plant expression system. The plant expression system provides for post-translational modification and processing to produce a recombinant gene product exhibiting enzymatic activity. The invention is demonstrated by working examples in which transgenic tobacco plants express recombinant expression constructs comprising human glucocerebrosidase nucleotide sequences. The invention is also demonstrated by working examples in which transfected tobacco plants express recombinant viral expression constructs comprising human &#x3b1; galactosidase nucleotide sequences. The recombinant lysosomal enzymes produced in accordance with the invention may be used for a variety of purposes, including but not limited to enzyme replacement therapy for the therapeutic treatment of human and animal lysosomal storage diseases.

Production of lysosomal enzymes in plants by transient expression

The present invention features a method for isolating and purifying proteins and peptides of interest from a plant host, which is applicable on a larger scale. Moreover, the present invention provides a more efficient method for isolating proteins and peptides of interest than those methods described in the prior art. In general, the present method of isolating proteins and peptides of interest comprises the steps of homogenizing a plant to produce a green juice, adjusting the pH of and heating the green juice, separating the target protein/peptide from other components of the green juice by one or more cycles of centrifugation and/or resuspension, and finally purifying proteins and peptides by such procedures as chromatography and/or salt precipitation.

Process for isolating and purifying proteins and peptides from plant sources

The present invention relates to foreign peptide sequences fused to recombinant plant viral structural proteins and a method of their production. Fusion proteins are economically synthesized in plants at high levels by biologically contained tobamoviruses. The fusion proteins of the invention have are useful as antigens for inducing the production of antibodies having desired binding properties, e.g., protective antibodies, or for use as vaccine antigens for the induction of protective immunity against the parvovirus. Feline parvovirus epitopes were fused to the N-terminus of the TMV coat protein, expressed in Nicotiana plants, extracted, purified, characterized and administered to animals, resulting in protective immunity.

Parvovirus vaccine as viral coat protein fusions

The present invention relates to a recombinant viral nucleic acid selected from a (&plus;) sense, single stranded RNA virus possessing a native subgenomic promoter encoding for a first viral subgenomic promoter, a nucleic acid sequence that codes for a viral coat protein whose transcription is regulated by the first viral subgenomic promoter, a second viral subgenomic promoter and a second nucleic acid sequence whose transcription is regulated by the second viral subgenomic promoter. The first and second viral subgenomic promoters of the recombinant viral nucleic acid do not have homologous sequences relative to each other. The recombinant viral nucleic acid provides the particular advantage that it systemically transcribes the second nucleic acid in the host. Host organisms encompassed by the present invention include procaryotes and eucaryotes, particularly animals and plants. The present invention also relates to viruses containing the viral vectors which are infective, production cells which are capable of producing the viruses or parts thereof, a host infected by the viruses of the invention, the gene products produced by expression of the viral nucleic acids and a process for the production of a desired product by growing the infected hosts.

Recombinant animal viral nucleic acids

The present invention features a method for isolating and purifying vitamins and sugars from a plant host which is applicable on a large scale. Moreover, the present invention provides a more efficient method for isolating vitamins and sugars than those methods described in the prior art. In general, the present method of isolating vitamins and sugars comprises the steps of homogenizing a plant to produce a green juice, adjusting the pH of and heating the green juice, separating the target species, either vitamins or sugars, from other components of the green juice by one or more cycles of centrifugation, resuspension, and ultrafiltration, and finally purifying vitamins or sugars by such procedure as PEG-precipitation, chromatography and/or salt precipitation.

Process for isolating and purifying viruses and sugars from plant sources

Growing community of inventors

Vacaville, CA, United States of America

Stephen J Garger