Vhh based binding antibodies for anthrax and botulinum toxins and methods of making and using therefor
AssigneeTufts University
InventorCHARLES B. SHOEMAKER
Life of this patent
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Abstract
Methods, compositions and kits are provided for treating a subject exposed to or at risk for exposure to a disease agent, methods, compositions and kits having a pharmaceutical composition including at least one recombinant binding protein or a source of expression of the binding protein, wherein the binding protein neutralizes at least one or a plurality of disease agents that are toxins, for example at least one of a Botulinum toxin or an Anthrax toxin.
Claims21
-25. (canceled)
A pharmaceutical composition for treating, ameliorating, or preventing disease or infection, or a symptom thereof, in a subject who has been exposed to, or who is at risk of, exposure to a Bacillus anthracis toxin, the pharmaceutical composition comprising a recombinant binding protein that specifically binds to and neutralizes the toxin, wherein the binding protein comprises an amino acid sequence selected from SEQ ID NO:
- 1, SEQ ID NO:
- 3, SEQ ID NO:
- 5, SEQ ID NO:7, SEQ ID NO:
- 9, SEQ ID NO:
- 11, SEQ ID NO:
- 13, SEQ ID NO:
- 15, SEQ ID NO:
- 105, SEQ ID NO:
- 106, SEQ ID NO:
- 107, SEQ ID NO:
- 108, SEQ ID NO:109, SEQ ID NO:
- 110, SEQ ID NO:
- 111, SEQ ID NO:
- 112, SEQ ID NO:
- 113, SEQ ID NO:
- 114, SEQ ID NO:
- 115, or a combination thereof.
A pharmaceutical composition for treating, ameliorating, or preventing disease or infection, or a symptom thereof, in a subject who has been exposed to, or who is at risk of, exposure to a Botulinum toxin B, the pharmaceutical composition comprising a recombinant binding protein that specifically binds to and neutralizes the toxin, wherein the binding protein comprises an amino acid sequence selected from SEQ ID NO:
- 131, SEQ ID NO:
- 133, SEQ ID NO:
- 135, SEQ ID NO:
- 137, or a combination thereof.
A pharmaceutical composition for treating, ameliorating, or preventing disease or infection, or a symptom thereof, in a subject who has been exposed to, or who is at risk of, exposure to a Botulinum toxin E, the pharmaceutical composition comprising a recombinant binding protein that specifically binds to and neutralizes the toxin, wherein the binding protein comprises an amino acid sequence selected from SEQ ID NO:
- 49, SEQ ID NO:
- 51, SEQ ID NO:
- 53, SEQ ID NO:
- 55, SEQ ID NO:
- 57, SEQ ID NO:
- 59, SEQ ID NO:
- 61, SEQ ID NO:
- 63, SEQ ID NO:
- 65, SEQ ID NO:
- 67, SEQ ID NO:
- 69, SEQ ID NO:
- 71, SEQ ID NO:
- 73, SEQ ID NO:
- 75, SEQ ID NO:
- 77, SEQ ID NO:79, SEQ ID NO:
- 81, SEQ ID NO:
- 83, SEQ ID NO:
- 85, SEQ ID NO:
- 87, SEQ ID NO:
- 89, SEQ ID NO:
- 91, SEQ ID NO:
- 93, SEQ ID NO:
- 95, SEQ ID NO:
- 97, SEQ ID NO:
- 99, SEQ ID NO:
- 101, SEQ ID NO:
- 127, SEQ ID NO:
- 129, SEQ ID NO:
- 139, SEQ ID NO:
- 141, SEQ ID NO:
- 143, or a combination thereof.
The pharmaceutical composition according to claim 1, wherein the B. anthracis toxin binding protein binds to and neutralizes at least one of an Anthrax surface generated protective antigen, an Anthrax lethal toxin, or an Anthrax edema toxin of B. anthracis.
The pharmaceutical composition according to claim 1, wherein the binding protein is heteromultimeric and comprises a plurality of non-identical binding regions, wherein each binding region specifically binds to and neutralizes a non-overlapping portion of the disease-causing agent.
The pharmaceutical composition according to claim 1, wherein the binding protein further comprises at least one of a tag epitope that is specifically bound by an anti-tag antibody and a linker that separates binding regions of the binding protein, wherein the linker comprises at least one of a peptide, a protein, a sugar, or a nucleic acid.
The pharmaceutical composition according to claim 2, wherein the binding protein is heteromultimeric and comprises a plurality of non-identical binding regions, wherein each binding region specifically binds to and neutralizes a non-overlapping portion of the disease-causing agent.
The pharmaceutical composition according to claim 2, wherein the binding protein further comprises at least one of a tag epitope that is specifically bound by an anti-tag antibody and a linker that separates binding regions of the binding protein, wherein the linker comprises at least one of a peptide, a protein, a sugar, or a nucleic acid.
The pharmaceutical composition according to claim 3, wherein the binding protein is heteromultimeric and comprises a plurality of non-identical binding regions, wherein each binding region specifically binds to and neutralizes a non-overlapping portion of the disease-causing agent.
The pharmaceutical composition according to claim 3, wherein the binding protein further comprises at least one of a tag epitope that is specifically bound by an anti-tag antibody and a linker that separates binding regions of the binding protein, wherein the linker comprises at least one of a peptide, a protein, a sugar, or a nucleic acid.
A pharmaceutical composition comprising one or more of the following:
- (i) a recombinant binding protein that specifically binds to and neutralizes a Bacillus anthracis toxin, wherein the binding protein comprises an amino acid sequence that has at least 90% identity to one or more sequences as set forth in SEQ ID NO:
- 1, SEQ ID NO:
- 3, SEQ ID NO:
- 5, SEQ ID NO:7, SEQ ID NO:
- 9, SEQ ID NO:
- 11, SEQ ID NO:
- 13, SEQ ID NO:
- 15, SEQ ID NO:
- 105, SEQ ID NO:
- 106, SEQ ID NO:
- 107, SEQ ID NO:
- 108, SEQ ID NO:109, SEQ ID NO:
- 110, SEQ ID NO:
- 111, SEQ ID NO:
- 112, SEQ ID NO:
- 113, SEQ ID NO:
- 114, or SEQ ID NO:
- 115;
- (ii) a recombinant binding protein that specifically binds to and neutralizes a Botulinum toxin B, wherein the binding protein comprises an amino acid sequence that has at least 90% identity to one or more sequences as set forth in SEQ ID NO:
- 131, SEQ ID NO:
- 133, SEQ ID NO:
- 135, or SEQ ID NO:
- 137;
- or
- (iii) a recombinant binding protein that specifically binds to and neutralizes a Botulinum toxin E, wherein the binding protein comprises an amino acid sequence that has at least 90% identity to one or more sequences as set forth in SEQ ID NO:
- 49, SEQ ID NO:
- 51, SEQ ID NO:
- 53, SEQ ID NO:
- 55, SEQ ID NO:
- 57, SEQ ID NO:
- 59, SEQ ID NO:
- 61, SEQ ID NO:
- 63, SEQ ID NO:
- 65, SEQ ID NO:
- 67, SEQ ID NO:
- 69, SEQ ID NO:
- 71, SEQ ID NO:
- 73, SEQ ID NO:
- 75, SEQ ID NO:
- 77, SEQ ID NO:79, SEQ ID NO:
- 81, SEQ ID NO:
- 83, SEQ ID NO:
- 85, SEQ ID NO:
- 87, SEQ ID NO:
- 89, SEQ ID NO:
- 91, SEQ ID NO:
- 93, SEQ ID NO:
- 95, SEQ ID NO:
- 97, SEQ ID NO:
- 99, SEQ ID NO:
- 101, SEQ ID NO:
- 127, SEQ ID NO:
- 129, SEQ ID NO:
- 139, SEQ ID NO:
- 141, or SEQ ID NO:
- 143.
A method of detecting or identifying a bacterial toxin in a sample, the method comprising:
- contacting the sample with at least one detectably labeled binding protein that specifically binds the bacterial toxin, wherein the binding protein comprises an amino acid sequence selected from one or more of SEQ ID NO:
- 1, SEQ ID NO:
- 3, SEQ ID NO:
- 5, SEQ ID NO:7, SEQ ID NO:
- 9, SEQ ID NO:
- 11, SEQ ID NO:
- 13, SEQ ID NO:
- 15, SEQ ID NO:
- 17, SEQ ID NO:
- 19, SEQ ID NO:
- 21, SEQ ID NO:
- 23, SEQ ID NO:
- 25, SEQ ID NO:
- 27, SEQ ID NO:
- 29, SEQ ID NO:
- 31, SEQ ID NO:
- 33, SEQ ID NO:
- 35, SEQ ID NO:
- 37, SEQ ID NO:
- 39, SEQ ID NO:
- 41, SEQ ID NO:
- 43, SEQ ID NO:
- 45, SEQ ID NO:
- 47, SEQ ID NO:
- 49, SEQ ID NO:
- 51, SEQ ID NO:
- 53, SEQ ID NO:
- 55, SEQ ID NO:
- 57, SEQ ID NO:
- 59, SEQ ID NO:
- 61, SEQ ID NO:
- 63, SEQ ID NO:
- 65, SEQ ID NO:
- 67, SEQ ID NO:
- 69, SEQ ID NO:
- 71, SEQ ID NO:
- 73, SEQ ID NO:
- 75, SEQ ID NO:
- 77, SEQ ID NO:79, SEQ ID NO:
- 81, SEQ ID NO:
- 83, SEQ ID NO:
- 85, SEQ ID NO:
- 87, SEQ ID NO:
- 89, SEQ ID NO:
- 91, SEQ ID NO:
- 93, SEQ ID NO:
- 95, SEQ ID NO:
- 97, SEQ ID NO:
- 99, SEQ ID NO:
- 101, SEQ ID NO:
- 103, SEQ ID NO:
- 105, SEQ ID NO:
- 107, SEQ ID NO:
- 109, SEQ ID NO:
- 111, SEQ ID NO:
- 113, SEQ ID NO:
- 115, SEQ ID NO:
- 117, SEQ ID NO:
- 119, SEQ ID NO:
- 121, SEQ ID NO:
- 123, SEQ ID NO:
- 125, SEQ ID NO:
- 127, SEQ ID NO:
- 129, SEQ ID NO:
- 131, SEQ ID NO:
- 133, SEQ ID NO:
- 135, SEQ ID NO:
- 137, SEQ ID NO:
- 139, SEQ ID NO:
- 141, or SEQ ID NO:
- 143 under conditions for the binding protein to interact with the bacterial toxin;
- and
- measuring the level of binding of the binding protein to the bacterial toxin in the sample relative to a control to detect or identify the presence of the toxin in the sample.
A method for treating a subject who has been exposed to or who is at risk for exposure to disease or infection by a bacterial toxin, the method comprising:
- administering to the subject a polynucleotide which encodes a binding protein comprising one or more binding regions that specifically bind and neutralize a bacterial toxin selected from at least one of Bacillus anthracis toxin, Botulinum toxin B, or Botulinum toxin E, wherein the binding protein comprises an amino acid sequence selected from SEQ ID NO:
- 1, SEQ ID NO:
- 3, SEQ ID NO:
- 5, SEQ ID NO:7, SEQ ID NO:
- 9, SEQ ID NO:
- 11, SEQ ID NO:
- 13, SEQ ID NO:
- 15, SEQ ID NO:
- 17, SEQ ID NO:
- 19, SEQ ID NO:
- 21, SEQ ID NO:
- 23, SEQ ID NO:
- 25, SEQ ID NO:
- 27, SEQ ID NO:
- 29, SEQ ID NO:
- 31, SEQ ID NO:
- 33, SEQ ID NO:
- 35, SEQ ID NO:
- 37, SEQ ID NO:
- 39, SEQ ID NO:
- 41, SEQ ID NO:
- 43, SEQ ID NO:
- 45, SEQ ID NO:
- 47, SEQ ID NO:
- 49, SEQ ID NO:
- 51, SEQ ID NO:
- 53, SEQ ID NO:
- 55, SEQ ID NO:
- 57, SEQ ID NO:
- 59, SEQ ID NO:
- 61, SEQ ID NO:
- 63, SEQ ID NO:
- 65, SEQ ID NO:
- 67, SEQ ID NO:
- 69, SEQ ID NO:
- 71, SEQ ID NO:
- 73, SEQ ID NO:
- 75, SEQ ID NO:
- 77, SEQ ID NO:79, SEQ ID NO:
- 81, SEQ ID NO:
- 83, SEQ ID NO:
- 85, SEQ ID NO:
- 87, SEQ ID NO:
- 89, SEQ ID NO:
- 91, SEQ ID NO:
- 93, SEQ ID NO:
- 95, SEQ ID NO:
- 97, SEQ ID NO:
- 99, SEQ ID NO:
- 101, SEQ ID NO:
- 103, SEQ ID NO:
- 105, SEQ ID NO:
- 107, SEQ ID NO:
- 109, SEQ ID NO:
- 111, SEQ ID NO:
- 113, SEQ ID NO:
- 115, SEQ ID NO:
- 117, SEQ ID NO:
- 119, SEQ ID NO:
- 121, SEQ ID NO:
- 123, SEQ ID NO:
- 125, SEQ ID NO:
- 127, SEQ ID NO:
- 129, SEQ ID NO:
- 131, SEQ ID NO:
- 133, SEQ ID NO:
- 135, SEQ ID NO:
- 137, SEQ ID NO:
- 139, SEQ ID NO:
- 141, SEQ ID NO:
- 143, or a combination thereof, thereby treating disease or infection in the subject.
A method of treating, ameliorating, or preventing infection or disease, or a symptom thereof, in a subject who has been exposed to or who is at risk of exposure to a Bacillus anthracis toxin, a Botulinum toxin B, and/or a Botulinum toxin E, the method comprising:
- administering to a subject in need thereof one or more of the following:
- (i) a recombinant binding protein that specifically binds to and neutralizes a Bacillus anthracis toxin, wherein the binding protein comprises an amino acid sequence that has at least 90% identity to one or more sequences as set forth in SEQ ID NO:
- 1, SEQ ID NO:
- 3, SEQ ID NO:
- 5, SEQ ID NO:7, SEQ ID NO:
- 9, SEQ ID NO:
- 11, SEQ ID NO:
- 13, SEQ ID NO:
- 15, SEQ ID NO:
- 105, SEQ ID NO:
- 106, SEQ ID NO:
- 107, SEQ ID NO:
- 108, SEQ ID NO:109, SEQ ID NO:
- 110, SEQ ID NO:
- 111, SEQ ID NO:
- 112, SEQ ID NO:
- 113, SEQ ID NO:
- 114, or SEQ ID NO:
- 115;
- (ii) a recombinant binding protein that specifically binds to and neutralizes a Botulinum toxin B, wherein the binding protein comprises an amino acid sequence that has at least 90% identity to one or more sequences as set forth in SEQ ID NO:
- 131, SEQ ID NO:
- 133, SEQ ID NO:
- 135, or SEQ ID NO:
- 137;
- or
- (iii) a recombinant binding protein that specifically binds to and neutralizes a Botulinum toxin E, wherein the binding protein comprises an amino acid sequence that has at least 90% identity to one or more sequences as set forth in SEQ ID NO:
- 49, SEQ ID NO:
- 51, SEQ ID NO:
- 53, SEQ ID NO:
- 55, SEQ ID NO:
- 57, SEQ ID NO:
- 59, SEQ ID NO:
- 61, SEQ ID NO:
- 63, SEQ ID NO:
- 65, SEQ ID NO:
- 67, SEQ ID NO:
- 69, SEQ ID NO:
- 71, SEQ ID NO:
- 73, SEQ ID NO:
- 75, SEQ ID NO:
- 77, SEQ ID NO:79, SEQ ID NO:
- 81, SEQ ID NO:
- 83, SEQ ID NO:
- 85, SEQ ID NO:
- 87, SEQ ID NO:
- 89, SEQ ID NO:
- 91, SEQ ID NO:
- 93, SEQ ID NO:
- 95, SEQ ID NO:
- 97, SEQ ID NO:
- 99, SEQ ID NO:
- 101, SEQ ID NO:
- 127, SEQ ID NO:
- 129, SEQ ID NO:
- 139, SEQ ID NO:
- 141, or SEQ ID NO:
- 143, thereby treating, ameliorating, or preventing infection or disease, or a symptom thereof.
Dependencies are inferred from the claim text.