Methods, compositions and kits for treating a subject using a recombinant neutralizing binding protein
Methods, compositions and kits are provided for treating a subject exposed to or at risk for exposure to a disease agent using a pharmaceutical composition including at least one recombinant binding protein or a source of expression of the binding protein, wherein the binding protein neutralizes a disease agent that is a toxin, for example a Clostridium difficile toxin, a Shiga toxin, a ricin toxin, or an anthrax toxin.
1. An isolated recombinant binding protein comprising an amino acid sequence selected from SEQ ID NO: 100, SEQ ID NO: 102, SEQ ID NO: 104, SEQ ID NO: 106, SEQ ID NO: 108, SEQ ID NO: 110, SEQ ID NO: 112, SEQ ID NO: 114, SEQ ID NO: 116, SEQ ID NO: 118, SEQ ID NO: 120, or SEQ ID NO: 122, which specifically binds to Anthrax protective antigen (PA) toxin.
2. A composition comprising at least one recombinant binding protein that specifically binds to Anthrax protective antigen (PA) toxin, wherein the at least one binding protein comprises an amino acid sequence selected from SEQ ID NO: 100, SEQ ID NO: 102, SEQ ID NO: 104, SEQ ID NO: 106, SEQ ID NO: 108, SEQ ID NO: 110, SEQ ID NO: 112, SEQ ID NO: 114, SEQ ID NO: 116, SEQ ID NO: 118, SEQ ID NO: 120, or SEQ ID NO: 122.
3. A composition comprising at least one recombinant binding protein that specifically binds to Anthrax protective antigen (PA) toxin, wherein the at least one binding protein is encoded by a polynucleotide sequence comprising a nucleotide sequence selected from 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, or SEQ ID NO: 123.
4. The composition of claim 2 , further comprising a pharmaceutically acceptable carrier, excipient, or vehicle.
5. A multimeric binding protein comprising at least two recombinant camelid heavy-chain-only antibody monomers (VHHs) which specifically bind Anthrax protective antigen (PA) toxin and which comprise an amino acid sequence selected from SEQ ID NO: 100, SEQ ID NO: 102, SEQ ID NO: 104, SEQ ID NO: 106, SEQ ID NO: 108, SEQ ID NO: 110, SEQ ID NO: 112, SEQ ID NO: 114, SEQ ID NO: 116, SEQ ID NO: 118, SEQ ID NO: 120, or SEQ ID NO: 122; wherein the at least two VHHs are separated by spacers.
6. The multimeric binding protein of claim 5 , comprising two VHHs which bind Anthrax PA toxin.
7. The multimeric binding protein of claim 5 , further comprising one or more epitope tags to which an anti-epitope tag antibody specifically binds.
8. The multimeric binding protein of claim 5 , wherein the spacer comprises a peptide having an amino acid sequence selected from GGGGS (SEQ ID NO: 54), GGGGSGGGGSGGGGS (SEQ ID NO: 55), or a combination thereof.
9. The multimeric binding protein of claim 7 , wherein the one or more epitope tags comprises the amino acid sequence Gly Ala Pro Val Pro Tyr Pro Asp Pro Leu Glu Pro Arg of SEQ ID NO: 15.
10. A pharmaceutical composition comprising the multimeric binding protein of claim 5 and a pharmaceutically acceptable carrier, excipient, or vehicle.
11. A method of targeting and binding to Anthrax protective antigen (PA) toxin, said method comprising contacting said PA toxin with the composition of claim 4 in an amount effective to target and bind to said Anthrax PA toxin.
12. The method according to claim 11 , wherein the Anthrax PA toxin is targeted and bound in vitro or in vivo.
13. A method of treating or preventing intoxication of a subject by Anthrax protective antigen (PA) toxin, said method comprising: administering to a subject in need thereof the composition of claim 4 , in an amount effective for the binding protein to bind to and neutralize Anthrax PA toxin in the subject after intoxication or prior to the subject's having symptoms of intoxication.