Methods, compositions and kits for treating a subject using a recombinant neutralizing binding protein

1. A pharmaceutical composition comprising a recombinant binding protein that binds to and neutralizes at least two different disease agents, wherein the binding protein comprises at least four disease agent binding domains separated by spacer peptides; wherein the disease agent binding domains comprise the amino acid sequences

(SEQ ID NO: 174)

QVQLVESGGGLVQPGGSLRLSCAASGFTLDYSSIGWFRQAPGKEREGVSC

ISSSGDSTKYADSVKGRFTTSRDNAKNTVYLQMNSLKPDDTAVYYCAAFR

ATMCGVFPLSPYGKDDWGKGTLVTVSSEPKTPKPQ;

(SEQ ID NO: 164)

QLQLVESGGGLVQPGGSLRLSCEASGFTLDYYGIGWFRQPPGKEREAVSY

ISASARTILYADSVKGRFTISRDNAKNAVYLQMNSLKREDTAVYYCARRR

FSASSVNRWLADDYDVWGRGTQVAVSSEPKTPKPQ;

(SEQ ID NO: 165)

QVQLVESGGGLVQTGGSLRLSCASSGSIAGFETVTWSRQAPGKSLQWVAS

MTKTNNEIYSDSVKGRFIISRDNAKNTVYLQMNSLKPEDTGVYFCKGPEL

RGQGIQVTVSSEPKTPKPQ;

and

(SEQ ID NO: 166)

QVQLVESGGGLVQPGGSLRLSCAASGFTFSDYVMTWVRQAPGKGPEWIAT

INTDGSTMRDDSTKGRFTISRDNAKNTLYLQMTSLKPEDTALYYCARGRV

ISASAIRGAVRGPGTQVTVSSEPKTPKPQ.

2. The pharmaceutical composition according to claim 1 , wherein one of the disease agents bound and neutralized by the recombinant binding protein is a toxin produced by C. difficile which causes C. difficile infection (CDI) or a C. difficile -associated disease selected from C. difficile diarrhea (CDD), antibiotic-associated diarrhea, C. difficile intestinal inflammatory disease, colitis, pseudomembranous colitis, peritonitis, megacolon, colon perforation and rupture and symptoms thereof.

3. The pharmaceutical composition according to claim 1 , wherein each of the disease agent binding domains is separated from another disease agent binding domain by a flexible spacer peptide amino acid sequence.

4. The pharmaceutical composition according to claim 3 , wherein the flexible spacer peptide amino acid sequence comprises amino acid sequence GGGGSGGGGSGGGGS (SEQ ID NO: 55).

5. The pharmaceutical composition according to claim 1 , wherein at least one of the disease agent binding domains binds and neutralizes toxin A of Clostridium difficile (TcdA).

6. The pharmaceutical composition according to claim 1 , wherein at least one of the disease agent binding domains binds and neutralizes toxin B of Clostridium difficile (TcdB).

7. The pharmaceutical composition according to claim 1 , wherein the pharmaceutical composition comprises one or more of the amino acids sequences of SEQ ID NOs: 167, 171, 172, and 173.

8. The pharmaceutical composition according to claim 1 , wherein the disease agent binding domains bind and neutralize toxin A of Clostridium difficile (TcdA) and toxin B of Clostridium difficile (TcdB).

9. A method of treating or preventing an antibiotic-induced adverse effect in the GI tract associated with C. difficile infection, comprising administering to a subject in need thereof an effective amount of the pharmaceutical composition according to claim 1 .

10. The method according to claim 9 , wherein the subject is receiving or likely to receive one or more antibiotic treatments, optionally selected from fluoroquinolones, cephalosporins, clindamycin and penicillins.

11. The method according to claim 9 , wherein the pharmaceutical composition further comprises one or more of SEQ ID NO: 174, SEQ ID NO: 164, SEQ ID NO: 165, SEQ ID NO: 166, SEQ ID NO: 167, SEQ ID NO: 171, SEQ ID NO: 172, and SEQ ID NO: 173.

12. A method of treating or preventing a C. difficile infection (CDI) or C. difficile -associated disease, the method comprising administering to a subject in need thereof an effective amount of the pharmaceutical composition according to claim 1 .

13. The method according to claim 12 , wherein the C. difficile infection (CDI) or C. difficile -associated disease is one or more of: antibiotic-associated diarrhea, C. difficile diarrhea (CDD), C. difficile intestinal inflammatory disease, colitis, pseudomembranous colitis, fever, abdominal pain, dehydration and disturbances in electrolytes, megacolon, peritonitis, and perforation and rupture of the colon.