Method of preparing conjugated antibodies

Claim 1

A method of preparing a conjugated antibody comprising (i) attaching a substantially pure antigen to a solid phase; (ii) passing an antibody containing composition over said solid phase such that said antibody binds to said attached antigen to form a bound antigen-antibody complex; (iii) passing a reactive chelating agent over said bound antigen-antibody complex to chelating said conjugating agent to said complex; and (iv) removing the resultant conjugated antibody from said solid phase.

Claim 2

A method of preparing a conjugated antibody comprising (i) attaching a substantially pure antigen to a solid phase; (ii) passing an antibody containing composition over said solid phase such that said antibody binds to said attached antigen to form a bound antigen-antibody complex; (iii) passing a reactive chelating agent over said bound antigen-antibody complex to chelating said conjugating agent to said complex; and (iv) removing the resultant conjugated antibody from said solid phase.

Claim 3

A method of preparing a conjugated antibody comprising: (i) reacting a substantially pure antigen with an antibody to form a bond antigen-antibody complex; (ii) conjugating a reactive chelating agent with said bound antigen-antibody complex; and (iii) cleaving said antigen from said antibody to form a conjugated antibody.

Claim 4

A method of preparing a conjugated antibody comprising: (i) reacting a substantially pure antigen with an antibody to form a bond antigen-antibody complex; (ii) conjugating a reactive chelating agent with said bound antigen-antibody complex; and (iii) cleaving said antigen from said antibody to form a conjugated antibody.

Claim 5

A method of preparing a conjugated antibody comprising the steps of (I) (a) packing a solid phase in the column of a syringe; (b) washing said solid phase with 0.001M HCl; (c) coupling calcitonin HCO 3 - buffer for two hours at room temperature; (d) washing the column with HCO 3 - buffer; (e) reacting said beads with 1M ethanolamine at about pH 8; (f) washing said beads with 0.1M acetate buffer at about pH 4; (g) washing said beads with 0.1M borate buffer at about pH 8; (h) repeating said steps (f) and (g); (i) washing said column with an affinity antibody containing solution; (j) washing said column with phosphate buffered saline to form an immuno-adsorbent bead complex with said antibody; (II) conjugating said immuno-adsorbent bead complex with a chelating agent to form a conjugated antibody by (a) washing said column with an HCO 3 - buffer; (b) flushing said column with diethylamine triamine pentacetic acid anhydride; (c) washing said column with phosphate buffered saline; (d) eluting said antibody chelate conjugate with 6M guanidine HCl; and (III) separating said antibody conjugate from said guanidine HCl by gel filtration.

Claim 6

A method of preparing a conjugated antibody comprising the steps of (I) (a) packing a solid phase in the column of a syringe; (b) washing said solid phase with 0.001M HCl; (c) coupling calcitonin HCO 3 - buffer for two hours at room temperature; (d) washing the column with HCO 3 - buffer; (e) reacting said beads with 1M ethanolamine at about pH 8; (f) washing said beads with 0.1M acetate buffer at about pH 4; (g) washing said beads with 0.1M borate buffer at about pH 8; (h) repeating said steps (f) and (g); (i) washing said column with an affinity antibody containing solution; (j) washing said column with phosphate buffered saline to form an immuno-adsorbent bead complex with said antibody; (II) conjugating said immuno-adsorbent bead complex with a chelating agent to form a conjugated antibody by (a) washing said column with an HCO 3 - buffer; (b) flushing said column with diethylamine triamine pentacetic acid anhydride; (c) washing said column with phosphate buffered saline; (d) eluting said antibody chelate conjugate with 6M guanidine HCl; and (III) separating said antibody conjugate from said guanidine HCl by gel filtration.