Patent ID: 11965882
Assignee: CELL.COPEDIA GMBH
Field: Measurement (Instruments)
Classification: CPC G  C | IPC C  G

Claim 0:
1. A method of isolating a biological entity from a sample comprising:
(i) providing a sample comprising the biological entity (1), wherein the biological entity comprises a surface antigen (2);
(ii) providing a first tagging agent (3) comprising at least one binding domain (4) and a ligand (5), wherein the binding domain (4) is capable of specifically binding to the surface antigen (2) on the biological entity via a first antigen recognition interaction (9);
(iii) providing a carrier (11) comprising a first ligand binding partner (7) and a second ligand binding partner (7), wherein the first ligand binding partner (7) is capable of forming a first non-covalent protein-ligand interaction (8) with the ligand (5) of the first tagging agent (3);
(iv) providing a second tagging agent (3) comprising a ligand (5) and at least one binding domain (4), wherein the ligand (5) is capable of forming a second non-covalent protein-ligand interaction (8) with the second ligand binding partner (7) of the carrier (11);
(v) providing a linking molecule (10) comprising an antigen (2′) and a ligand (5), wherein the antigen (2′) is capable of specifically binding to the binding domain (4) of the second tagging agent (3) via a second antigen recognition interaction (9); and
(vi) providing freely movable beads (6), wherein the beads (6) are non-magnetic beads, and wherein the beads comprise a third ligand binding partner (7) capable of forming a third non-covalent protein-ligand interaction (8) with the ligand (5) of the linking molecule (10);
(vii) incubating the sample, the first tagging agent (3), the carrier (11), the second tagging agent (3), the linking molecule (10) and the beads (6) within a container and allowing complex formation between the biological entity (1), the first tagging agent (3) specifically bound to the biological entity (1) via the first antigen recognition interaction (9), the carrier (11) immobilizing the first tagging agent (3) via the first non-covalent protein-ligand interaction (8), the second tagging agent (3) immobilized on the carrier (11) via the second non-covalent protein-ligand interaction (8), the linking molecule (10) specifically bound to the second tagging agent (3) via the second antigen recognition interaction (9), and the beads (6) immobilizing the linking molecule (10) via the third non-covalent protein-ligand interaction (8); and
(viii) purifying the biological entity (1) by temporarily holding the beads (6) in place in the container, while discarding the supernatant, and adding a washing buffer; and/or
(ix) isolating the biological entity (1) by releasing the biological entity (1) from the beads (6) and the tagging agent (3), respectively, and recovery of the biological entity (1) from the supernatant while temporarily holding the beads (6) in place in the container, optionally allowing for the recovery of the first and second tagging agents (3) and/or beads (6).