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Human Blood Proteins Expressed in Monocot Seeds

a monocot seed and protein technology, applied in the field of human blood proteins produced, can solve the problems of short or limited supply of human blood proteins, and none of these patents or publications discloses the production of human blood proteins in monocot seeds in high yield

Inactive Publication Date: 2010-02-04
INVITRIA INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Many human blood proteins are in short or limited supply due to the larger quantities required of the protein for positive therapeutic effect or possibly also due to the larger demand of these proteins by the world population of patients having the particular condition.
None of these patents or publications discloses the production of human blood proteins in monocot seeds in high yield.

Method used

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  • Human Blood Proteins Expressed in Monocot Seeds
  • Human Blood Proteins Expressed in Monocot Seeds
  • Human Blood Proteins Expressed in Monocot Seeds

Examples

Experimental program
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Effect test

example 1

Production of Transgenic Rice Encoding AAT and Fibrinogen Polypeptides

[0106]The basic procedures of particle bombardment-mediated rice transformation and plant regeneration were carried out as described by Huang et al., 2001. Rice variety TP309 seeds were dehusked, sterilized in 50% (v / v) commercial bleach for 25 min and washed with sterile water. The sterilized seeds were placed on rice callus induction medium (RCI) plates containing [N6 salts (Sigma), B5 vitamins (Sigma), 2 mg / l 2,4-D and 3% sucrose]. The rice seeds were incubated for 10 days to induce callus formation. Primary callus was dissected from the seeds and placed on RCI for 3 weeks. This was done twice more to generate secondary and tertiary callus which was used for bombardment and continued subculture. A callus of 1-4 mm diameter was placed in a 4 cm circle on RCI with 0.3M mannitol 0.3M sorbitol for 5-24 hrs prior to bombardment. Microprojectile bombardment was carried out using the Biolistic PDC-1000 / He system (Bio-...

example 2

Production of Rice Extract Containing Recombinant Blood Proteins and its Use in Parenteral and Enteric Formulations

General Procedure for Production of Rice Extract

[0108]Transgenic rice containing heterologous polypeptides can be converted to rice extracts by either a dry milling or wet milling process. In the dry milling process, transgenic paddy rice seeds containing the heterologous polypeptides were dehusked with a dehusker. The rice was grounded into a fine flour though a dry milling process, for example, in one experiment, at speed 3 of a model 91 Kitchen Mill from K-TEC. Phosphate buffered saline (“PBS”), containing 0.135 N NaCl, 2.7 mM KCl, 10 mM Na2HPO4, 1.7 mM KH2PO4, at pH 7.4, with or without additional NaCl, such as 0.35 N NaCl, was added to the rice flour. In some experiments, approximately 10 ml of extraction buffer was used for each 1 g of flour. In other experiments, the initial flour / buffer ratio varied over a range such as 1 g / 40 ml to 1 g / 10 ml. The mixture was in...

example 3

Concentration and Diafiltration of Recombinant Blood Protein and Control Rice Extracts

[0116]The conditions used in concentration and diafiltration vary depending on volume, speed, cost, etc. These conditions are standard in the art based on the description herein. The frozen initial extract was thawed in the coldroom (about 2-8° C.) for six hours. The thawed material was clarified though a 0.45 μm filter and concentrated using a 5000 Nominal Molecular Weight Cutoff membrane of polyethersulfone.

[0117]90 ml of the filtrate of control extract was concentrated to 10 ml and additional 10 ml of deionized water can be added to the concentrated filtrate. The diluted filtrate can be diafiltrated one more time using water. The precipitate starts forming at 16 mS and increases as the ionic strength decreases. A solution of 1.0M ammonium bicarbonate was added to the retentate to add ionic strength. The haze decreases although does not disappear completely. The material was diafiltered multiple ...

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PUM

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Abstract

The invention is directed to blood proteins produced in monocot seeds and isolated therefrom for use in therapeutic compositions, and to methods of making these isolated blood proteins and to therapeutic compositions comprising them.

Description

[0001]This application is a continuation of pending U.S. patent application Ser. No. 10 / 411,395, filed Apr. 11, 2003, which in turn is a continuation-in-part of U.S. application Ser. No. 10 / 077,381, filed Feb. 14, 2002, which claims priority benefit to U.S. provisional application Ser. No. 60 / 269,199, filed Feb. 14, 2001, application Ser. No. 10 / 077,381 being a continuation-in-part of U.S. patent application Ser. No. 09 / 847,232, filed May 2, 2001, which claims priority benefit to U.S. provisional application Ser. No. 60 / 266,929, filed Feb. 6, 2001, and U.S. provisional application Ser. No. 60 / 201,182, filed May 2, 2000. All priority applications are incorporated herein by reference in their entirety.FIELD OF THE INVENTION[0002]The present invention relates to human blood proteins produced in the seeds of monocot plants for use in making human and animal topical compositions and human therapeutic compositions.BACKGROUND OF THE INVENTION[0003]Many human blood proteins are in short or ...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): C12N15/82A01H5/10A23K1/14A23K1/16A23L1/30A23L1/305A23L7/10A23L7/20A23L29/238C07K14/415C07K14/485C07K14/65C07K14/79C07K14/81C12N9/08C12N9/36G01N33/574
CPCA23K1/14A23K1/1631G01N2500/02A23L1/1041A23L1/185A23L1/3002A23L1/3056A23V2002/00B82Y30/00C07K14/415C07K14/485C07K14/65C07K14/79C07K14/8125C12N9/0065C12N9/2462C12N15/8216C12N15/8222C12N15/8234C12N15/8257G01N33/57484G01N33/57492G01N2333/4725A23V2300/21A23K10/30A23K20/147A23L7/198A23L7/20A23L33/105A23L33/19A61P3/02A61P7/00A61P7/02A61P7/04A61P7/06
Inventor HUANG, NINGRODRIQUEZ, RAYMOND L.HAGIE, FRANK E.STALKER, DAVID M.
Owner INVITRIA INC
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