US 12,304,947 B2
Recombinant antibody against human cardiac troponin I
Peng Cui, Dongguan (CN); Zhiqiang He, Dongguan (CN); Yuan Meng, Dongguan (CN); Dongmei Zhong, Dongguan (CN); Hui You, Dongguan (CN); and Lingyun Fan, Dongguan (CN)
Assigned to Fapon Biotech Inc., Shenzhen (CN)
Appl. No. 17/284,234
Filed by FAPON BIOTECH INC., Shenzhen (CN)
PCT Filed Sep. 27, 2019, PCT No. PCT/CN2019/108682
§ 371(c)(1), (2) Date Apr. 9, 2021,
PCT Pub. No. WO2020/073832, PCT Pub. Date Apr. 16, 2020.
Claims priority of application No. 201811179512.3 (CN), filed on Oct. 10, 2018.
Prior Publication US 2021/0340231 A1, Nov. 4, 2021
Int. Cl. C07K 16/18 (2006.01); G01N 33/68 (2006.01)
CPC C07K 16/18 (2013.01) [G01N 33/6893 (2013.01); C07K 2317/565 (2013.01); C07K 2317/92 (2013.01); G01N 2800/12 (2013.01); G01N 2800/324 (2013.01)] 20 Claims
 
1. An isolated binding protein comprising an antigen-binding domain, wherein the antigen-binding domain comprises six complementarity determining regions of the following amino acid sequences, and has an affinity for cardiac troponin I:
a complementarity determining region CDR-VH1 having a sequence of G-F-N-X1-K-X2-Y-X3-M-H (SEQ ID NO: 23), where
X1 is L or I, X2 is D, and X3 is F or Y;
a complementarity determining region CDR-VH2 having a sequence of R-I-X1-P-E-D-X2-E-T-X3-Y-A-P-E (SEQ ID NO: 24), where X1 is D, X2 is A or G, and X3 is R or K;
a complementarity determining region CDR-VH3 having a sequence of Y-Y-X1-S-Y-X2-P-F-V-Y (SEQ ID NO:25), where
X1 is S, and X2 is I, L or V;
a complementarity determining region CDR-VL1 having a sequence of Q-S-X1-X2-Y-S-N-X3-H-T-Y (SEQ ID NO:26), where
X1 is I or L, X2 is I or L, and X3 is R;
a complementarity determining region CDR-VL2 having a sequence of Q-X1-S-X2-R-F-S(SEQ ID NO: 27), where X1 is I, L or V, and X2 is N; and
a complementarity determining region CDR-VL3 having a sequence of S-X1-S-T-H-X2-P-X3-T (SEQ ID NO: 28), where
X1 is Q, X2 is L or I, and X3 is F or Y, and
wherein the mutation site of each of the complementarity determining regions is selected from any one of the following mutation combinations:
 
 
 CDR- CDR- CDR- CDR- CDR- CDR-
 
 VH1 VH2 VH3 VL1 VL2 VL3
Site X1/X3 X2/X3 X2 X1/X2 X1 X2/X3
 
Mutation L/F A/R I I/I I L/F
combination 1 
 
 
 
 
 
 
Mutation L/Y A/K L I/L L L/Y
combination 2 
 
 
 
 
 
 
Mutation I/F G/R V L/I V I/F
combination 3 
 
 
 
 
 
 
Mutation I/Y G/K L L/L L I/Y
combination 4 
 
 
 
 
 
 
Mutation L/F A/K V L/L V L/F
combination 5 
 
 
 
 
 
 
Mutation L/Y G/R I I/I I I/F
combination 6 
 
 
 
 
 
 
Mutation I/F G/K V I/L V I/Y
combination 7 
 
 
 
 
 
 
Mutation I/Y A/R I L/I I L/Y
combination 8 
 
 
 
 
 
 
Mutation L/F G/R L L/I L L/F
combination 9 
 
 
 
 
 
 
Mutation L/Y G/K I L/L I I/Y
combination 10
 
 
 
 
 
 
Mutation I/F A/R L I/I L L/Y
combination 11
 
 
 
 
 
 
Mutation I/Y A/K V I/L V I/F
combination 12
 
 
 
 
 
 
Mutation L/F G/K L I/L L L/Y
combination 13
 
 
 
 
 
 
Mutation L/Y A/R V L/I V L/F
combination 14
 
 
 
 
 
 
Mutation I/F A/K I L/L I I/F
combination 15
 
 
 
 
 
 
Mutation I/Y G/R V I/I V I/Y
combination 16
 
 
 
 
 
 
Mutation L/F A/R I L/L I L/Y
combination 17
 
 
 
 
 
 
Mutation L/Y A/R L I/I L I/Y
combination 18
 
 
 
 
 
 
Mutation I/F A/K I I/L I L/F
combination 19
 
 
 
 
 
 
Mutation I/Y G/R L L/I L I/F
combination 20
 
 
 
 
 
 
Mutation L/F G/K V LIT V I/F
combination 21
 
 
 
 
 
 
Mutation L/Y A/K L L/L L L/F
combination 22
 
 
 
 
 
 
Mutation I/F G/R V I/I V L/Y
combination 23
 
 
 
 
 
 
Mutation I/Y G/K I I/L I I/Y
combination 24
 
 
 
 
 
 
Mutation L/F A/R V L/L V I/F
combination 25
 
 
 
 
 
 
Mutation L/Y G/R I I/I I L/Y
combination 26
 
 
 
 
 
 
Mutation I/F G/K L I/L L L/F
combination 27
 
 
 
 
 
 
Mutation I/Y A/R I L/I I I/Y
combination 28
 
 
 
 
 
 
Mutation L/F A/K L L/I L L/F
combination 29
 
 
 
 
 
 
Mutation L/Y G/K V L/L V L/Y
combination 30
 
 
 
 
 
 
Mutation I/F A/R L I/I L I/F
combination 31
 
 
 
 
 
 
Mutation I/Y A/K V I/L V I/Y
combination 32
 
 
 
 
 
 
Mutation L/F G/R I I/L I L/F
combination 33
 
 
 
 
 
 
Mutation L/Y A/K V L/I V I/F
combination 34
 
 
 
 
 
 
Mutation I/F G/K I L/L I I/Y
combination 35
 
 
 
 
 
 
Mutation I/Y A/R L I/I L L/Y
combination 36
 
 
 
 
 
 
Mutation L/Y G/R L I/I L I/Y
combination 37
 
 
 
 
 
 
Mutation I/F G/K V I/L V L/Y
combination 38
 
 
 
 
 
 
Mutation I/Y A/K I L/I I I/F
combination 39
 
 
 
 
 
 
Mutation L/F G/R V LIT V L/Y
combination 40
 
 
 
 
 
 
Mutation L/Y G/K I L/L I L/F
combination 41
 
 
 
 
 
 
Mutation I/Y G/R I I/L I I/Y
combination 42
 
 
 
 
 
 
Mutation L/F G/K L L/L L L/Y
combination 43
 
 
 
 
 
 
Mutation L/Y A/R V I/I V I/Y
combination 44
 
 
 
 
 
 
Mutation I/F A/K L I/L L L/F
combination 45
 
 
 
 
 
 
Mutation I/Y G/K V L/I V I/F
combination 46
 
 
 
 
 
 
Mutation L/F A/R I L/I I I/F
combination 47
 
 
 
 
 
 
Mutation L/Y A/K V L/L V L/F
combination 48
 
 
 
 
 
 
Mutation I/F G/R I I/I I L/Y
combination 49
 
 
 
 
 
 
Mutation I/Y A/R L I/L L I/Y
combination 50
 
 
 
 
 
 
Mutation L/F A/K I I/L I I/F
combination 51
 
 
 
 
 
 
Mutation L/Y G/R L L/I L L/Y
combination 52
 
 
 
 
 
 
Mutation I/F A/R V L/L V L/F
combination 53
 
 
 
 
 
 
Mutation I/Y A/K L I/I L  I/Y.
combination 54