Production and uses of type i ribosome inactivating proteins
AssigneeBioo Scientific Corporation
InventorMingang Chen
Life of this patent
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Abstract
Described herein are methods to express and purify recombinant type 1 ribosome inactivating proteins. Included are methods for using recombinant cucurmosin as a therapeutic to treat cancer and infectious diseases.
Claims50
A method for preparing substantially purified recombinant type 1 ribosome inactivating protein (RIP) comprising:
- obtaining a protein sequence comprising at least a portion of a recombinant type 1 RIP;
- synthesizing a nucleic acid encoding the type 1 RIP using a gene sequence to express the type 1 RIP;
- cloning the nucleic acid into an appropriate expression vector;
- transferring the expression vector to an appropriate protein expression system;
- inducing the expression of the recombinant type 1 RIP;
- and
- substantially purifying the expressed recombinant type 1 RIP.
A pharmaceutical composition suitable for use in the treatment of cancer, viral infection, or any other disease in human and non human organisms said pharmaceutical composition comprising an effective amount of natural or recombinant cucurmosin, or functional derivatives thereof in combination with an appropriate pharmaceutical carrier medium.
A derivative of recombinant cucurmosin, said derivative comprising one or more of a ligand conjugated to cucurmosin, a truncated cucurmosin polypeptide, or a cucurmosin polypeptide comprising at least one mutation in the polypeptide sequence thereof.
A method of inducing apoptosis in cancer cells comprising contacting cancer cells with an at least partially purified preparation of recombinant cucurmosin, or a functional derivative thereof.
A method for preparing substantially purified recombinant cucurmosin comprising:
- obtaining a nucleic acid sequence comprising at least a portion of cucurmosin;
- cloning the nucleic acid into the appropriate expression vector;
- transferring the expression vector to an appropriate protein expression system;
- inducing the expression of the recombinant cucurmosin;
- and
- substantially purifying the expressed recombinant cucurmosin.
A recombinant type 1 ribosome inactivating protein having the structure corresponding to SEQUENCE ID 5 and any modified derivative thereof.
An amino acid sequence having the structure corresponding to SEQUENCE ID 4 and any modified derivative thereof.
An amino acid sequence having the structure corresponding to SEQUENCE ID 2 and any modified derivative thereof.
An nucleic acid sequence having the structure corresponding to SEQUENCE ID 1 and any modified derivative thereof.
An nucleic acid sequence having the structure corresponding to SEQUENCE ID 3 and any modified derivative thereof.
A method for preparing substantially purified recombinant type 1 RIP comprising:
- obtaining a nucleic acid sequence corresponding to SEQUENCE ID 1;
- cloning the nucleic acid into the appropriate expression vector;
- transferring the expression vector to an appropriate protein expression system;
- inducing the expression of the recombinant type 1 RIP;
- and
- substantially purifying the expressed recombinant type 1 RIP.
A method of purifying a tagged recombinant type 1 RIP, comprising:
- Expressing the type 1 RIP;
- purifying the tagged recombinant type 1 RIP using an affinity column specific for the tag;
A purified tagged type 1 RIP for use as a therapeutic agent.
A purified type 1 RIP for use as a therapeutic agent
A therapeutic use of type I RIP in cancer treatment, viral treatment and bacterial treatment in any type of plant, animal, or insect.
A conjugate for the delivery of RIP to cells comprising:
- RIP;
- a ligand, wherein the ligand is capable of binding to a surface of a cell;
- and
- wherein RIP and the ligand are covalently coupled to each other.
A conjugate for the delivery of RIP to cells comprising:
- RIP;
- a ligand, wherein the ligand is capable of binding to a surface of a cell;
- and
- wherein RIP and the ligand are a fusion protein.
Dependencies are inferred from the claim text.