Granted Patent
B2
US 12,466,875 · App. 17/833,941 · Granted Nov 11, 2025
Supramolecular high affinity protein-binding system for purification of biomacromolecules
Inventors:
Honggang Cui (Lutherville, MD); Yi Li (Baltimore, MD); Xuankuo Xu (Boxborough, MA); Lye Lin Lock (Maynard, MA); Zhengjian Li (Sudbury, MA)
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Abstract
In certain embodiments, the present invention provides novel antibody purification methods and systems using a potentially simple and cost-efficient means. In some embodiments, customized Z-33 derived from Staphylococcus aureus Protein A is used to construct immuno-amphiphile molecules which can assemble into immunofibers in aqueous solution with bioactive epitopes on the surface and have IgG binding ability.
Claims (19)
1 . A method for purifying an antibody or an Fc fusion protein, the method comprising the steps of:
a) dissolving an immuno-amphiphile in an aqueous solution at a physiological pH to provide a dissolved immuno-amphiphile; and aging the dissolved immuno-amphiphile overnight to provide immunofibers (IFs);
b) mixing a sample containing an antibody or an Fc fusion protein with the IFs, and allowing the IFs to bind the Fc portion of the antibody or Fc fusion protein to form an immunofiber-antibody complex or immunofiber-Fc fusion protein complex in a solution;
c) separating the immunofiber-antibody complex or the immunofiber-Fc fusion protein complex from the solution by adding salt and centrifugation; and
d) dissociating the IFs from the antibody or Fc fusion protein and collecting the unbound antibody or Fc fusion protein,
wherein the immuno-amphiphile comprises an antibody binding peptide and a hydrocarbon moiety;
wherein the hydrocarbon moiety is conjugated to the N-terminus of the antibody binding peptide;
wherein the antibody binding peptide consists of the amino acid sequence FNMQQQRRFYEALHDPNLNEEQRNAKIKSIRDD (SEQ ID NO: 1); and
wherein the hydrocarbon moiety is a C12 hydrocarbon chain or two C8 hydrocarbon chains.
2 . The method of claim 1 , wherein the IFs are separated from the antibody or Fc fusion protein by lowering the pH to elution condition.
3 . The method of claim 1 , wherein the antibody binding peptide has an α-helical conformation when in an aqueous solution at physiological pH.
4 . The method of claim 1 , further comprising pretreating the immuno amphiphile in hexafluoroisopropanol prior to dissolving the immuno-amphiphile in the aqueous solution.
5 . The method of claim 4 , further comprising removing the hexafluoroisopropanol.
6 . The method of claim 5 , wherein removing the hexafluoroisopropanol comprises evaporating the hexafluoroisopropanol.
7 . The method of claim 1 , wherein the antibody is an IgG.
8 . The method of claim 1 , wherein the antibody is a monoclonal antibody.
9 . The method of claim 1 , wherein the IFs are separated from the antibody or Fc fusion protein by lowering the pH to elution condition and filtration or microfiltration.
10 . The method of claim 1 , wherein the physiological pH is 7.4.
11 . The method of claim 2 , wherein lowering the pH to elution condition comprises lowering the pH to 2.8.
Assignments (2)
ASSIGNMENT OF ASSIGNOR'S INTEREST
Recorded Aug 3, 2022
From: LI, ZHENJIAN; XU, XUANKUO; LOCK, LYE LIN
To: BRISTOL-MYERS SQUIBB COMPANY
Reel/Frame 060704/0679 →
ASSIGNMENT OF ASSIGNOR'S INTEREST
Recorded Aug 3, 2022
From: CUI, HONGGANG; LI, YI
To: THE JOHNS HOPKINS UNIVERSITY
Reel/Frame 060704/0695 →
Continuity (3)
Continuation
16498675
Provisional Application
62478886
· Mar 30, 2017
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