Protein modification of living cells using sortase
Non-genetically engineered mammalian cells modified by sortase-mediated conjugation of an agent thereto are provided. Methods of conjugating agents to nongenetically engineered mammalian cells using sortase are provided. Methods of using the cells, e.g. a method of modulating an immune response of a subject to an entity of interest, a method of neutralizing a substance in the body of a subject, a method of treating a subject in need of treatment for deficiency of a protein, and a method of treating a subject in need of treatment for a disease, are provided.
1. A human cell having a binding moiety or therapeutic agent linked thereto via a sortase recognition sequence, wherein the binding moiety or therapeutic agent is linked to an endogenous, non-genetically engineered protein expressed by the human cell, and wherein the human cell has not been genetically engineered to express a polypeptide comprising a sortase recognition sequence, wherein the binding moiety or therapeutic agent comprises an immune checkpoint inhibitor.
2. The human cell of claim 1 , wherein the human cell is an immune system cell.
3. The human cell of claim 1 , wherein the sortase recognition sequence comprises LPXTG.
4. The human cell of claim 1 , wherein the binding moiety comprises a targeting moiety that binds to an epitope or antigen of interest.
5. The human cell of claim 1 , wherein the binding moiety comprises a monoclonal antibody, a single chain antibody, or a single domain antibody.
6. The human cell of claim 5 , wherein the binding moiety comprises a VHH.
7. The human cell of claim 1 , wherein the human cell is selected from the group consisting of a red blood cell, a peripheral blood mononuclear cell, a lymphocyte, a T cell, a NK cell, and a dendritic cell.
8. The human cell of claim 1 , wherein the human cell is a non-immortalized cell.
9. The human cell of claim 1 , wherein the human cell is a primary cell.
10. The human cell of claim 1 , wherein the human cell has not been genetically engineered to express a polypeptide comprising a sequence comprising one or more N-terminal glycines.
11. The human cell of claim 1 , wherein the human cell has not been genetically engineered.
12. A method of treating a subject in need of treatment for a disease, the method comprising administering at least one human cell of claim 1 to the subject, wherein the at least one cell is effective for treating the disease.
13. The method of claim 12 , wherein the human cell originates from the subject and said subject is in need of evaluation or treatment for a disease of interest or from a donor who is immunocompatible with the subject.
14. The method of claim 13 , wherein the human cell originates from the subject in need of evaluation or treatment for a disease characterized by the presence of abnormal or excessive cells or pathogens in the subject's body, or from a donor who is immunocompatible with the subject.
15. The method of claim 14 , wherein the human cell originates from the subject in need of evaluation or treatment for cancer, an autoimmune disease, or an infection or from a donor who is immunocompatible with the subject.
16. A mammalian cell having a binding moiety or therapeutic agent linked thereto via a sortase recognition sequence, wherein the binding moiety or therapeutic agent is linked to an endogenous, non-genetically engineered protein expressed by the mammalian cell, and wherein the binding moiety or therapeutic agent comprises an immune checkpoint inhibitor.
17. A method of treating a subject in need of treatment for a disease, the method comprising administering the mammalian cell of claim 16 to the subject.