Transgenic plants expressing assembled secretory antibodies

• US 5,959,177 A
• Filed: 05/03/1996
• Issued: 09/28/1999
• Est. Priority Date: 10/27/1989
• Status: Expired due to Fees

• Current Assignee: The Scripps Research Institute
• Original Assignee: The Scripps Research Institute
• Inventors: Hiatt, Andrew, Hein, Mich B., Ma, Julian K-C
• Primary Examiner(s): Robinson, Douglas W.
• Assistant Examiner(s): HAAS, THOMAS M
• Application Number:
  US08/642,406
• Time in Patent Office: 1,243 Days
• Field of Search: 800/209, 435/172.3, 435/320.1, 435/419, 435/69.1, 536/23.5, 536/25.53, 536/24.1
• US Class Current: 800/288
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: C07K 16/00 Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies antibodies with enzymatic activity, e.g. abzymes C12N9/0002
• CPC Class Codes: C07K 16/12 against material from bacteria
• CPC Class Codes: C07K 16/1228 from Enterobacteriaceae (F), e.g. Citrobacter, Serratia, Proteus, Providencia, Morganella, Yersinia
• CPC Class Codes: C07K 16/1275 from Streptococcus (G)
• CPC Class Codes: C07K 16/44 against material not provided for elsewhere , e.g. haptens, metals, DNA, RNA, amino acids
• CPC Class Codes: C07K 2317/13 isolated from plants
• CPC Class Codes: C07K 2319/00 Fusion polypeptide
• CPC Class Codes: C07K 2319/02 containing a signal sequence
• CPC Class Codes: C12N 15/8242 with non-agronomic quality (output) traits, e.g. for industrial processing; Value added, non-agronomic traits
• CPC Class Codes: C12N 15/8258 for the production of oral vaccines (antigens) or immunoglobulins
• CPC Class Codes: C12N 9/0002 Antibodies with enzymatic activity, e.g. abzymes
• CPC Class Codes: C12N 9/18 Carboxylic ester hydrolases (3.1.1)

Abstract

The present invention relates to expression and assembly of foreign multimeric proteins--e.g., antibodies--in plants, as well as to transgenic plants that express such proteins. In one of several preferred embodiments, the generation and assembly of functional secretory antibodies in plants is disclosed. The invention also discloses compositions produced by the transgenic plants of the present invention and methods of using same.

Claims

1. A method of generating and assembling secretory antibodies within a single cell, said method comprising:

a. introducing into the genome of a first member of a plant species a first mammalian nucleotide sequence encoding an immunoglobulin alpha heavy chain portion-containing polypeptide including a leader sequence forming a secretion signal, to produce a first transformant;

b. introducing into the genome of a second member of said plant species a second mammalian nucleotide sequence encoding a polypeptide linker or joining chain, to produce a second transformant;

c. introducing into the genome of a third member of said plant species a third mammalian nucleotide sequence encoding a secretory component, to produce a third transformant;

d. sexually crossing said transformants to generate a progeny population containing all three mammalian sequences; and

e. isolating from said progeny population a transgenic plant species producing a secretory antibody.

2. The method of claim 1, wherein said method further comprises introducing into the genome of a fourth member of said plant species a fourth mammalian nucleotide sequence encoding an immunoglobulin light chain portion-containing polypeptide including a leader sequence forming a secretion signal, to produce a fourth transformant;

sexually crossing said fourth transformants with said other transformants to generate a progeny population containing all four mammalian sequences; and

isolating from said progeny population a transgenic plant species producing a secretory antibody.

3. The method of claim 1, wherein said first mammalian nucleotide sequence encodes a single-chain antibody.

4. The method of claim 1, wherein said first mammalian nucleotide sequence encodes an immunoglobulin alpha heavy chain portion-containing polypeptide including more than one variable region.

5. The method of claim 1, wherein nucleotide sequences are introduced via separate vectors.

6. A transgenic plant comprising:

a. plant cells that contain a nucleotide sequence encoding one or more immunoglobulin heavy-chain polypeptides, a nucleotide sequence encoding a polypeptide linker or joining chain, and a nucleotide sequence encoding a secretory component; and

b. immunologically active secretory antibodies encoded by said nucleotide sequences.

7. The plant of claim 6, further comprising a nucleotide sequence encoding one or more light-chain polypeptides.

8. The plant of claim 6, wherein all three nucleotide sequences are contained within a single cell.

9. The plant of claim 7, wherein all four nucleotide sequences are contained within a single cell.

10. The plant of claim 6, which is a dicotyledonous plant.

11. The plant of claim 6, which is a monocotyledonous plant.

12. The plant of claim 6, wherein each of said nucleotide sequences is included on a separate vector.