Polymerization and copolymerization of proteins
First Claim
1. A method for producing a polyprotein having at least 10 units held together by sulfur to sulfur or sulfur to carbon bonds, each unit comprising a protein and one or more heterobifunctional reagents capable of forming a covalent bond with an amino group wherein at least one of the heterobifunctional reagents is capable of forming a covalent sulfur-sulfur or carbon-sulfur bond with a thiol group and at least one of the reagents has a thiol group or a group capable of being converted into a thiol group, the method comprising the steps of:
- a) forming a first protein-heterobifunctional reagent complex by treating a first protein with a sufficient amount of a heterobifunctional reagent so as to form a covalent bond between amino groups of the first protein and 3 to 5 molecules of the reagent per protein molecule;
b) forming a second protein-heterobifunctional reagent complex by treating a second protein with a sufficient amount of a heterobifunctional reagent so as to form a covalent bond between amino groups of the second protein and at least 5 molecules of the reagent per protein unit;
c) providing a thiol group on at least one of the reagents; and
d) joining the first protein-heterobifunctional reagent complex and the second protein-heterobifunctional reagent complex by forming sulphur-sulphur or carbon-sulphur bonds into a polyprotein having at least 10 units.
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Abstract
A method for producing a polyprotein having at least 10 units, and often as many as 50 to 100 and more units held together by sulfur to sulfur or sulfur to carbon bonds is disclosed. Each unit comprises a protein and one or more heterobifunctional reagents. One functional group of the reagent is capable of forming a covalent bond with an amino group, permitting the reagent to bind to a protein. The other functional group of the reagent is capable of forming a covalent bond with a thiol group so as to form the covalent sulfur-carbon or sulfur-sulfur bond with another heterobifunctional reagent bonded to another protein.
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Citations
48 Claims
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1. A method for producing a polyprotein having at least 10 units held together by sulfur to sulfur or sulfur to carbon bonds, each unit comprising a protein and one or more heterobifunctional reagents capable of forming a covalent bond with an amino group wherein at least one of the heterobifunctional reagents is capable of forming a covalent sulfur-sulfur or carbon-sulfur bond with a thiol group and at least one of the reagents has a thiol group or a group capable of being converted into a thiol group, the method comprising the steps of:
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a) forming a first protein-heterobifunctional reagent complex by treating a first protein with a sufficient amount of a heterobifunctional reagent so as to form a covalent bond between amino groups of the first protein and 3 to 5 molecules of the reagent per protein molecule; b) forming a second protein-heterobifunctional reagent complex by treating a second protein with a sufficient amount of a heterobifunctional reagent so as to form a covalent bond between amino groups of the second protein and at least 5 molecules of the reagent per protein unit; c) providing a thiol group on at least one of the reagents; and d) joining the first protein-heterobifunctional reagent complex and the second protein-heterobifunctional reagent complex by forming sulphur-sulphur or carbon-sulphur bonds into a polyprotein having at least 10 units. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 46, 47, 48)
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30. A polyprotein that has substantially the solubility properties of its component protein units, and that contains at least 10 protein units having molecular weight greater than 1×
- 104, said polyprotein is formed by a method comprising forming a first protein-heterobifunctional reagent complex, forming a second protein-heterobifunctional reagent complex, and joining said first protein-heterobifunctional reagent complex and said second protein-heterobifunctional reagent complex.
- View Dependent Claims (31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45)
Specification