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US 10,377,816 B2Granted
Nucleotide sequences, vectors and host cells
AssigneeGlaxosmithkline Biologicals SA
InventorCindy Castado
Life of this patent
Estimating the term…
Abstract
Polynucleotides encoding fusion proteins comprising fragments of toxin A and toxin B from Clostridium difficile are described, as well as vectors and host cells containing such polynucleotides.
Claims11
A polypeptide comprising a N-terminal first fragment of toxin A and a C-terminal second fragment of toxin B, wherein
- (i) the N-terminal first fragment is a Clostridium difficile toxin A repeating domain fragment and comprises at least 100 amino acids, wherein Clostridium difficile toxin A is represented by SEQ ID NO:1;
- and
- (ii) the C-terminal second fragment is a Clostridium difficile toxin B repeating domain fragment and comprises at least 100 amino acids, wherein Clostridium difficile toxin B is represented by SEQ ID NO:2;
- (iii) the N-terminal first fragment comprises a first proximal C-terminal end within a first repeat portion, said first repeat portion including a portion of repeat portion VII or repeat portion VIII of toxin A, wherein the first proximal C-terminal end is within the long repeat VII of toxin A (amino acids 2614-2644 of SEQ ID NO:
- 1) or the short repeat VIII of toxin A (amino acids 2645 to 2660);
- (iv) the C-terminal second fragment comprises a second proximal N-terminal end within a second repeat portion, said second repeat portion including a portion of repeat portion I of toxin B (amino acids 1897-1926 of SEQ ID NO:2) or repeat portion II of toxin B (amino acids 2028-2057 of SEQ ID NO:2);
- and
- wherein the first proximal C-terminal end of the N-terminal first fragment and the second proximal N-terminal end of the C-terminal second fragment are joined by 5 to 0 amino acids such that β-solenoid structure is maintained through the junction between the two fragments,
- wherein the polypeptide elicits antibodies that neutralize toxin A or toxin B or both,
- wherein:
- said first proximal C-terminal end of the N-terminal first fragment of toxin A is within a short repeat portion of toxin A and said second proximal N-terminal end of the second C-terminal fragment of toxin B is within a short repeat portion of toxin B, or
- said first proximal C-terminal end of the N-terminal first fragment of toxin A is within a long repeat portion of toxin A and said second proximal N-terminal end of the second C-terminal fragment of toxin B is within a long repeat portion of toxin B,
- wherein the first repeat portion and the second repeat portion have greater than 40% sequence identity to one another.
10 more elements
A polypeptide comprising a N-terminal first fragment of toxin A and a C-terminal second fragment of toxin B, wherein
- (i) the N-terminal first fragment is a Clostridium difficile toxin A repeating domain fragment and comprises at least 100 amino acids, wherein Clostridium difficile toxin A is represented by SEQ ID NO:1;
- and
- (ii) the C-terminal second fragment is a Clostridium difficile toxin B repeating domain fragment and comprises at least 100 amino acids, wherein Clostridium difficile toxin B is represented by SEQ ID NO:2;
- (iii) the N-terminal first fragment comprises a first proximal C-terminal end within a first repeat portion, said first repeat portion including a portion of repeat portion VII or repeat portion VIII of toxin A, wherein the first proximal C-terminal end is within an area selected from the long repeat VII of toxin A (amino acids 2614-2644 of SEQ ID NO:1) and amino acids 2620 to 2660 of toxin A;
- (iv) the C-terminal second fragment comprises a second proximal N-terminal end within a second repeat portion, said second repeat portion including a portion of repeat portion II of toxin B, wherein the second proximal N-terminal end is within the long repeat II of toxin B (amino acids 2028-2057 of SEQ ID NO:2);
- and
- wherein the first proximal C-terminal end of the N-terminal first fragment and the second proximal N-terminal end of the C-terminal second fragment are joined by 5 to 0 amino acids such that β-solenoid structure is maintained through the junction between the two fragments,
- wherein the polypeptide elicits antibodies that neutralize toxin A or toxin B or both,
- wherein:
- said first proximal C-terminal end of the N-terminal first fragment of toxin A is within a short repeat portion of toxin A and said second proximal N-terminal end of the second C-terminal fragment of toxin B is within a short repeat portion of toxin B, or
- said first proximal C-terminal end of the N-terminal first fragment of toxin A is within a long repeat portion of toxin A and said second proximal N-terminal end of the second C-terminal fragment of toxin B is within a long repeat portion of toxin B,
- wherein the first repeat portion and the second repeat portion have greater than 40% sequence identity to one another.
9 more elements
Dependencies are inferred from the claim text.