Methods for the production of multimeric immunoglobulins, and related compositions

Filing Information

• Publication Number: US20030093832
• Application Number: US10176380
• Filing date: 06/21/2002
• Publication date: 05/15/2003
• Predicted expiration date: 12/04/2021

• U.S. Classifications: 800/281  · 435/419  · 530/388.26  · 800/288  ·
• International Classifications: C07K016/40 · C12N005/04 · A01H005/00 ·
• Related U.S. Application Data:
  RELATED APPLICATIONS
  0001 This application is a continuation-in-part of U.S. utility application Ser. No. 10/032,201, filed on Dec. 19, 2001, which is a continuation-in-part of U.S. utility application Ser. No. 10/006,038, filed Dec. 4, 2001 to van Rooijen, et al., entitled “METHODS FOR THE PRODUCTION OF REDOX PROTEINS”; which is a continuation-in-part of U.S. utility application Ser. No. 09/742,900, filed Dec. 19, 2000 to Heifetz, et al., entitled “METHOD OF PRODUCTION AND DELIVERY OF THIOREDOXIN”. The Ser. 10/032,201 application is also a continuation-in-part of U.S. utility application Ser. No. 09/742,900 and claims benefit of priority under 35 U.S.C. §119(e) to U.S. provisional application Serial No. 60/302,885, filed Jul. 5, 2001, to van Rooijen, et al., entitled “METHODS FOR THE PRODUCTION OF REDOX PROTEINS”. The subject matter of each of the provisional and utility applications is incorporated herein by reference in its entirety.
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Patent Family

Abstract

Improved methods for the production of multimeric-protein-complexes, such as redox proteins and immunoglobulins, in association with oil bodies are described. The redox protein is enzymatically active when prepared in association with the oil bodies. Also provided are related nucleic acids, proteins, cells, plants, and compositions.

Independent Claims | See all claims (76)

1. 1. A method of producing an oil body associated with a recombinant multimeric-immunoglobulin, said method comprising: (a) producing in a cell comprising oil bodies, a first immunoglobulin-polypeptide-chain and a second immunoglobulin-polypeptide-chain wherein said first immunoglobulin-polypeptide-chain is capable of associating with said second immunoglobulin-polypeptide-chain to form said multimeric-immunoglobulin; and (b) associating said multimeric-immunoglobulin with an oil body through an oil-body-targeting-protein capable of associating with said oil body and said first immunoglobulin-polypeptide-chain.
2. 17. A method of expressing a recombinant multimeric-immunoglobulin comprising a first and second immunoglobulin-polypeptide-chain in a cell, said method comprising: (a) introducing into a cell a first chimeric nucleic acid sequence comprising: (i) a first nucleic acid sequence capable of regulating transcription in said cell operatively linked to; (ii) a second nucleic acid sequence encoding a first immunoglobulin-polypeptide-chain; (b) introducing into said cell a second chimeric nucleic acid sequence comprising: (i) a third nucleic acid sequence capable of regulating transcription in said cell operatively linked to, (ii) a fourth nucleic acid sequence encoding a second immunoglobulin-polypeptide-chain; (c) growing said cell under conditions to permit expression of said first and second immunoglobulin-polypeptide-chain in a progeny cell comprising oil bodies wherein said first recombinant immunoglobulin polypeptide-chain and said second immunoglublin-polypeptide-chain are capable of forming a multimeric-immunoglobulin; and (d) associating said first immunoglobulin-polypeptide-chain with an oil body through an oil-body-targeting-protein capable of associating With said oil body and said first immunoglobulin-polypeptide-chain.
3. 33. A method of producing in a plant a recombinant multimeric-immunoglobulin, said method comprising: (a) preparing a first plant comprising cells, said cells comprising oil bodies and a first immunoglobulin-polypeptide-chain wherein said first immunoglobulin-polypeptide-chain is capable of associating with said oil bodies through an oil-body-targeting-protein; (b) preparing a second plant comprising cells, said cells comprising oil bodies and a second immunoglobulin-polypeptide-chain; and (c) sexually crossing said first plant with said second plant to produce a progeny plant comprising cells, said cells comprising oil bodies, wherein said oil bodies are capable of associating with said first immunoglobulin-polypeptide-chain and said first recombinant immunoglobulin polypeptide is capable of associating with said second immunoglobulin-polypeptide-chain to form said recombinant multimeric-immunoglobulin.
4. 44. A method of producing in a plant a recombinant multimeric-immunoglobulin comprising: (a) preparing a first plant comprising cells, said cells comprising oil bodies and a first and second immunoglobulin-polypeptide-chain wherein said first immunoglobulin-polypeptide-chain is capable of associating with said oil bodies through an oil-body-targeting-protein; (b) preparing a second plant comprising cells, said cells comprising oil bodies and an oil-body-targeting-protein capable of associating with said first immunoglobulin-polypeptide-chain through said oil-body-targeting-protein; and (c) sexually crossing said first plant with said second plant to produce a progeny plant comprising cells, said cells comprising oil bodies, wherein said oil bodies are capable of associating with said first immunoglobulin-polypeptide-chain through an oil-body-targeting-protein, and said first immunoglobulin-polypeptide-chain is capable of associating with said second immunoglobulin-polypeptide-chain to form said recombinant multimeric-immunoglobulin.
5. 45. A method for preparing a multimeric-immunoglobulin associated with oil bodies comprising: a) introducing into a cell a chimeric nucleic acid sequence comprising: 1) a first nucleic acid sequence capable of regulating transcription in said cell operatively linked to; 2) a second nucleic acid sequence encoding a recombinant fusion polypeptide comprising (i) a nucleic acid sequence encoding a sufficient portion of an oil-body-protein to provide targeting of said recombinant fusion polypeptide to an oil body linked to (ii) a nucleic acid sequence encoding encoding a immunoglobulin comprising a first immunoglobulin-polypeptide-chain linked to a second immunoglobulin-polypeptide-chain, operatively linked to; 3) a third nucleic acid sequence capable of terminating transcription in said cell; b) growing said cell under conditions to permit expression of said multimeric-immunoglobulin in a progeny cell comprising oil bodies; and c) isolating from said progeny cell said oil bodies comprising said multimeric-immunoglobulin.
6. 46. A chimeric nucleic acid sequence comprising: 1) a first nucleic acid sequence capable of regulating transcription in a host cell operatively linked to; 2) a second nucleic acid sequence encoding a recombinant fusion polypeptide comprising (i) a nucleic acid sequence encoding a sufficient portion of an oil-body-protein to provide targeting of said recombinant fusion polypeptide to an oil body linked to (ii) a nucleic acid sequence encoding an immunoglobulin comprising a first immunoglobulin-polypeptide-chain linked to a second immunoglobulin-polypeptide-chain operatively linked to; 3) a third nucleic acid sequence capable of terminating transcription in said cell.
7. 47. A chimeric nucleic acid sequence encoding a multimeric-immunoglobulin-fusion-protein, said nucleic acid sequence comprising: (a) a first nucleic acid sequence encoding an oil-body-targeting-protein operatively linked in reading frame to; (b) a second nucleic acid sequence encoding a first immunoglobulin-polypeptide-chain; linked in reading frame to; (c) a third nucleic acid sequence encoding a second immunoglobulin-polypeptide-chain, wherein said first and second immunoglobulin-polypeptide-chain are capable of forming a multimeric-immunoglobulin.
8. 54. Isolated oil bodies comprising: (a) a first fusion protein comprising a first oil-body-targeting-protein fused to a first immunoglobulin-polypeptide-chain; and (b) a second fusion protein comprising a second oil-body-targeting-protein fused to a second-immunoglobulin-polypeptide-chain, wherein said first and second immunoglobulin-polypeptide-chain are capable of forming a multimeric-immunoglobulin.
9. 61. A cell comprising oil bodies and (i) an oil-body-targeting-protein, (ii) a first immunoglobulin-polypeptide-chain and (iii) a second immunoglobulin-polypeptide-chain wherein (1) said first immunoglobulin-polypeptide-chain is capable of associating with said oil-body-targeting-protein, and (2) said first immunoglobulin-polypeptide-chain capable of associating with said second immunoglobulin-polypeptide-chain to form a multimeric-immunoglobulin.