Methods and compositions of chemically modified phage libraries
Provided is a chemically modified phage display platform and method of use thereof. More specifically, the present disclosure provides a chemically modified phage display library that incorporates 2-acetylphenylboronic acid (APBA) moieties to elicit dynamic covalent binding to the bacterial cell surface. The APBA-modified phage display libraries described herein are applicable to a wide array of bacterial strains and/or mammalian cells, paving the way to facile diagnosis and development of strain-specific antibiotics, and/or peptide-antibiotic conjugates for effective and targeted treatment. Also provided are therapeutic peptides, and pharmaceutical compositions thereof, that are identified by screening the phage display library of the present disclosure, and method of use of such therapeutic peptides for effective and targeted treatment.
1. A phage display library comprising phage particles comprising phage displayed peptides comprising two acetylphenylboronic acid (APBA) modified cysteine residues.
2. The phage display library of claim 1 , wherein the phage displayed peptides comprise a peptide sequence given by:
XC*(X) n C*(X) m ,
wherein C* indicates an APBA modified cystein residue; wherein each instance of X is an amino acid independently selected from D, E, K, R, H, Y, N, Q, S, T, G, A, V, L, I, M, P, F, and W; wherein n is an integer selected from 5, 6, 7, 8, 9, and 10; wherein m is an integer selected from 1, 2, 3, 4, and 5.
3. The phage display library of claim 2 , wherein the phage display peptides comprise a peptide sequence given by:
AC*(X) n C*(G) m .
4. The phage display library of claim 2 , wherein n is 6, 7, or 8; and wherein m is 2, 3, or 4.
5. The phage display library of claim 1 , wherein the APBA modified cysteine residue has a structure given by a formula:
wherein each of A 1 and A 2 are independently a C1-C6 alkyl.
6. The phage display library of claim 5 , wherein the APBA modified cysteine residue has a structure given by a formula:
7. An acetylphenylboronic acid (APBA) peptide comprising a peptide sequence given by:
XC*(X) n C*(X) m ,
wherein C* indicates an APBA modified cystein residue; wherein each instance of X is an amino acid independently selected from D, E, K, R, H, Y, N, Q, S, T, G, A, V, L, I, M, P, F, and W; wherein n is an integer selected from 5, 6, 7, 8, 9, and 10; wherein m is an integer selected from 1, 2, 3, 4, and 5.
8. The APBA peptide of claim 7 , wherein the phage display peptides comprise a peptide sequence given by:
AC*(X) n C*(G) m .
9. The APBA peptide of claim 7 , wherein n is 6, 7, or 8; and wherein m is 2, 3, or 4.
10. The APBA peptide of claim 7 , wherein the APBA modified cysteine residue has a structure given by a formula:
wherein each of A 1 and A 2 are independently a C1-C6 alkyl.
11. The APBA peptide of claim 10 , wherein the APBA modified cysteine residue has a structure given by a formula:
12. The APBA peptide of claim 7 , further comprising an antibiotic residue, a phototoxin residue, or a detectable label residue at the N-terminus of the peptide.
13. The APBA peptide of claim 7 , further comprising an antibiotic residue, a phototoxin residue, or a detectable label residue at the C-terminus of the peptide.
14. A pharmaceutical composition comprising the APBA peptide of claim 7 and a pharmaceutically excipient.
15. The pharmaceutical composition of claim 14 , further comprising a therapeutic agent.
16. A drug screening method for selection of an APBA peptide, comprising contacting the phage display library of claim 1 to a target cell, and selecting a phage clone displaying a peptide capable of binding to the target cell.
17. The drug screening method of claim 16 , wherein the target cell is a bacterial cell.
18. The drug screening method of claim 17 , wherein the bacterial cell is a Staphylococcus aureus or Acinetobacter baumannii cell.
19. A method of developing a novel narrow-spectrum antibiotics for a bacterial strain of interest, comprising:
a) Screening the phage display library according to claim 1 with live bacteria of the bacterial strain of interest;
b) Selecting peptide binders with submicromolar affinity against the bacterial strain of interest;
c) Conjugating the selected peptide binders with an agent; and
d) Converting the peptide binders agent conjugate to an antibiotics targeting the bacterial strain of interest.
20. The method of claim 19 , wherein the bacterial strain is a Staphylococcus aureus or Acinetobacter baumannii strain.