USRE49495EActiveUtilityPatentIndex 59
Binding polypeptides having a mutated scaffold
Est. expiryAug 28, 2033(~7.1 yrs left)· nominal 20-yr term from priority
G01N 33/6845C07K 2319/00C07K 14/31C07K 1/047C12N 15/1044C07K 14/001C12Q 1/6874
59
PatentIndex Score
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Cited by
78
References
18
Claims
Abstract
The present disclosure relates to a class of engineered polypeptides and provides a polypeptide comprising the sequence EX2X3X4AX6X7EIX10 X11LPNLX16X17X18QX20 X21AFIX25X26LX28X29X30 PX32QSX35X36LLX39E AKKLX45X46X47Q (SEQ ID NO: 55). The present disclosure also relates to populations of polypeptide variants based on a common scaffold, each polypeptide in the population comprising the amino acid sequence EX2X3X4AX6X7EIX10 X11LPNLX16X17X18QX20 X21AFIX25X26LX28X29X30 PX32QSX35X36LLX39E AKKLX45X46X47Q (SEQ ID NO: 55), and methods for selecting a desired polypeptide having an affinity for a predetermined target from said population.
Claims
exact text as granted — not AI-modifiedThe invention claimed is:
1. A polypeptide comprising an amino acid sequence selected from:
i)
(SEQ ID NO: 55)
EX 2 X 3 X 4 AX 6 X 7 EIX 10 X 11 LPNLX 16 X 17 X 18 QX 20
X 21 AFIX 25 X 26 LX 28 X 29 X 30 PX 32 QSX 35 X 36 LLX 39 E
AKKLX 45 X 46 X 47 Q,
(SEQ ID NO: 55)
EX 2 X 3 X 4 AX 6 X 7 EIX 10 X 11 LPNLX 16 X 17 X 18 QX 20
X 21 AFIX 25 X 26 LX 28 X 29 X 30 PX 32 QSX 35 X 36 LLX 39 E
AKKLX 45 X 46 X 47 Q,
wherein each of X 2 , X 3 , X 4 , X 6 , X 7 , X 10 , X 11 , X 17 , X 18 , X 20 , X 21 , X 25 and X 28 independently corresponds to any amino acid residue; and
wherein, independently of each other,
X 16 is selected from N and T;
X 26 is selected from K and S;
X 29 X 30 PX 32 is selected from DDPS (SEQ ID NO: 74) and RQPE (SEQ ID NO: 75);
X 35 is selected from A and S;
X 36 is selected from E and N;
X 39 is selected from A, C and S;
X 45 is selected from E, N and S;
X 46 is selected from D, E and S, provided that X 46 is not D when X 45 is N;
X 47 is selected from A and S; and
ii) an amino acid sequence which has at least 91% identity to the sequence defined in i), provided that X 46 is not D or E when X 45 is N,
provided that for i) and ii) X 10 is not D when X 20 is W for a polypeptide capable of binding human complement component 5.
2. The polypeptide according to claim 1 , wherein X 45 is S.
3. The polypeptide according to claim 1 , wherein X 45 X 46 is selected from ES and SE.
4. The polypeptide according to claim 3 , wherein X 45 X 46 is SE.
5. The polypeptide according to claim 1 , which comprises an amino acid sequence selected from:
(SEQ ID NO: 56)
YAK EX 2 X 3 X 4 AX 6 X 7 EIX 10 X 11 LPNLX 16 X 17 X 18 QX 20
X 21 AFIX 25 X 26 LX 25 X 29 X 30 PX 32 QSX 35 X 36 LLX 39 E
AKKLX 45 X 46 X 47 Q AP;
and
(SEQ ID NO: 57)
FNK EX 2 X 3 X 4 AX 6 X 7 EIX 10 X 11 LPNLX 16 X 17 X 18 QX 20
X 21 AFIX 25 X 26 LX 25 X 29 X 30 PX 32 QSX 35 X 36 LLX 39 E
AKKLX 45 X 46 X 47 Q AP,
(SEQ ID NO: 56)
YAK EX 2 X 3 X 4 AX 6 X 7 EIX 10 X 11 LPNLX 16 X 17 X 18 QX 20
X 21 AFIX 25 X 26 LX 25 X 29 X 30 PX 32 QSX 35 X 36 LLX 39 E
AKKLX 45 X 46 X 47 Q AP;
and
(SEQ ID NO: 57)
FNK EX 2 X 3 X 4 AX 6 X 7 EIX 10 X 11 LPNLX 16 X 17 X 18 QX 20
X 21 AFIX 25 X 26 LX 25 X 29 X 30 PX 32 QSX 35 X 36 LLX 39 E
AKKLX 45 X 46 X 47 Q AP,
wherein each X y is as defined in claim 1 .
6. A fusion polypeptide comprising a polypeptide according to claim 1 as a moiety.
7. A polynucleotide encoding a polypeptide according to claim 1 or fusion polypeptide according to claim 6 .
8. A population of polypeptide variants based on a common scaffold, each polypeptide in the population comprising an amino acid sequence selected from:
i)
(SEQ ID NO: 55)
EX 2 X 3 X 4 AX 6 X 7 EIX 10 X 11 LPNLX 16 X 17 X 18 QX 20
X 21 AFIX 25 X 26 LX 28 X 29 X 30 PX 32 QSX 35 X 36 LLX 39 E
AKKLX 45 X 46 X 47 Q,
(SEQ ID NO: 55)
EX 2 X 3 X 4 AX 6 X 7 EIX 10 X 11 LPNLX 16 X 17 X 18 QX 20
X 21 AFIX 25 X 26 LX 28 X 29 X 30 PX 32 QSX 35 X 36 LLX 39 E
AKKLX 45 X 46 X 47 Q,
wherein each of X 2 , X 3 , X 4 , X 6 , X 7 , X 10 , X 11 , X 17 , X 18 , X 20 , X 21 , X 25 and X 28 independently corresponds to any amino acid residue; and
wherein, independently of each other,
X 16 is selected from N and T;
X 26 is selected from K and S;
X 29 X 30 PX 32 is selected from DDPS (SEQ ID NO: 74) and RQPE (SEQ ID NO: 75);
X 35 is selected from A and S;
X 36 is selected from E and N;
X 39 is selected from A, C and S;
X 45 is selected from E, N and S;
X 46 is selected from D, E and S, provided that X 46 is not D when X 45 is N;
X 47 is selected from A and S; and
ii) an amino acid sequence which has at least 91% identity to the sequence defined in i), provided that X 46 is not D or E when X 45 is N,
provided that for i) and ii) X 10 is not D when X 20 is W for a polypeptide capable of binding human complement component 5.
9. The population according to claim 8 , which comprises at least 1×10 4 unique polypeptide molecules.
10. A population of polynucleotides, characterized in that each member thereof encodes a member of a population of polypeptides according to claim 8 .
11. A composition comprising a polypeptide population according to claim 8 and a polynucleotide population according to claim 10 , wherein each member of said population of polypeptides is physically or spatially associated with the polynucleotide encoding that member via means for genotype-phenotype coupling.
12. The composition according to claim 11 , wherein said means for genotype-phenotype coupling comprises a phage display system.
13. A method for selecting a desired polypeptide having an affinity for a predetermined target from a population of polypeptides, comprising the steps:
(a) providing a population of polypeptides according to claim 8 ;
(b) bringing the population of polypeptides into contact with the predetermined target under conditions that enable specific interaction between the target and at least one desired polypeptide having an affinity for the target; and
(c) selecting, on the basis of said specific interaction, the at least one desired polypeptide from the remaining population of polypeptides.
14. A method for isolating a polynucleotide encoding a desired polypeptide having an affinity for a predetermined target, comprising the steps:
selecting said desired polypeptide and the polynucleotide encoding it from a population of polypeptides using the method according to claim 13 ; and
isolating the thus separated polynucleotide encoding the desired polypeptide.
15. A method for identifying a desired polypeptide having an affinity for a predetermined target, comprising the steps:
isolating a polynucleotide encoding said desired polypeptide using the method according to claim 14 ; and
sequencing the polynucleotide to establish by deduction the amino acid sequence of said desired polypeptide.
16. A method for selecting and identifying a desired polypeptide having an affinity for a predetermined target from a population of polypeptides, comprising the steps:
(a) synthesizing each member of a population of polypeptides according to claim 8 on a separate carrier or bead;
(b) selecting or enriching the carriers or beads based on the interaction of the polypeptide with the predetermined target; and
(c) identifying the polypeptide by protein characterization methodology.
17. The polypeptide according to claim 5, which comprises the amino acid sequence of SEQ ID NO: 56.
18. The polypeptide according to claim 5, which comprises the amino acid sequence of SEQ ID NO: 57.Cited by (0)
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