Supramolecular high affinity protein-binding system for purification of biomacromolecules
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.
1. An IgG binding immuno-amphiphile consisting of the amino acid sequence of SEQ ID NO: 1 conjugated at the N-terminus to a lauric acid moiety.
2. The IgG binding immuno-amphiphile of claim 1 , wherein a peptide portion of the IgG binding immuno-amphiphile has an α-helical conformation when in an aqueous solution at a pH between 2.8 to 7.4.
3. A conjugate of the IgG binding immuno-amphiphile of claim 1 with Rhodamine B.
4. A method for purification of an antibody or an Fc fusion protein, comprising the steps of:
dissolving the IgG binding immuno-amphiphile of claim 1 in an aqueous solution at physiological pH, and aging overnight to make it self-assemble into immunofibers (IFs);
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 and form an immunofiber-antibody complex or immunofiber-Fc fusion protein complex in solution;
separating the immunofiber-antibody complex or the immunofiber-Fc fusion protein complex from the solution; and
dissociating the IFs from the antibody or Fc fusion protein and collecting the unbound antibody or Fc fusion protein.
5. The method of claim 4 , wherein the IFs are separated from the antibody or Fc fusion protein by lowering the pH to elution condition and filtration or microfiltration.
6. The method of claim 4 , wherein the IFs are separated from the antibody or Fc fusion protein by salt-induced precipitation and centrifugation.
7. The method of claim 4 , wherein the IFs are separated from the antibody or Fc fusion protein by diafiltration.
8. The method of claim 4 , wherein a peptide portion of the IgG binding immuno-amphiphile has an α-helical conformation when in an aqueous solution at a pH between 2.8 to 7.4.
9. The method of claim 4 , wherein the IgG binding immuno-amphiphile is conjugated to Rhodamine B.