Methods for purifying heterodimeric multispecific antibodies from parental homodimeric antibody species
US-10787500-B2 · Sep 29, 2020 · US
US2023046406A1 · US · A1
| Field | Value |
|---|---|
| Publication number | US-2023046406-A1 |
| Application number | US-202117758779-A |
| Country | US |
| Kind code | A1 |
| Filing date | Feb 11, 2021 |
| Priority date | Mar 17, 2020 |
| Publication date | Feb 16, 2023 |
| Grant date | — |
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A method for separating of at least two transition metals, the method comprising: injecting a feed solution into a chromatography column comprising a chromatographic support media, the feed solution comprising at least two transition metals; eluting the feed from the column in an elution cycle by flowing an eluent through the column, wherein a concentration of the eluent is reduced during the elution cycle prior to elution of at least one of the transition metals.
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1 . A method for separating of at least two transition metals from each other, the method comprising: injecting a feed solution into a chromatography column comprising a chromatographic support media, the feed solution comprising at least two transition metals; eluting the feed from the column in an elution cycle by flowing an eluent through the column, wherein a concentration of the eluent is reduced during the elution cycle prior to elution of at least one of the transition metals. 2 . A method according to claim 1 , wherein one or more of the transition metals are selected from the group consisting of platinum group metals, gold, nickel, cobalt, manganese, copper, iron, rhenium, and technetium. 3 . A method according to claim 1 or 2 , wherein the feed solution comprises at least two platinum group metals and at least one base metal and/or amphoteric element and/or gold, and the method comprises separating the at least two platinum group metals from each other and from at least one base metal and/or at least one amphoteric element and/or gold, wherein the concentration of the eluent is reduced during the elution cycle prior to elution of at least one of the platinum group metals. 4 . A method according to any preceding claim, wherein the eluent is a halide salt or an acid halide. 5 . A method according to claim 4 , wherein the eluent is HCl. 6 . A method according to any preceding claims, wherein the concentration of eluent is reduced from at least 5 M to no more than 1 M. 7 . A method according to any preceding claims, wherein the concentration of the eluent is reduced as a step change from one concentration of eluent to a lower concentration of eluent. 8 . A method according to claim 7 , wherein the concentration of the eluent is further reduced by a second step change to a lower concentration of eluent during the elution cycle. 9 . A method according to any preceding claims, wherein the concentration of the eluent is increased prior to the end of the elution cycle. 10 . A method according to any preceding claims, wherein the feed comprises platinum and one or more of base metal, Rh, Ir, and Ru in a 3+ oxidation state, and wherein the concentration of the eluent is reduced after elution of the base metal, Rh, Ir, and/or Ru and prior to elution of the platinum from the column. 11 . A method according to claim 10 , wherein the feed further comprises palladium and the concentration of the eluent is reduced after elution of the base metal, Rh, Ir, and/or Ru and prior to or during the elution of the palladium. 12 . A method according to any one of claims 1 to 9 , wherein the feed comprises Rh and Ir in a 3+ oxidation state, and RuNO, and wherein the concentration of the eluent is reduced after elution of the Rh and Ir, and prior to elution of the RuNO from the column. 13 . A method according to any one of claims 1 to 9 , wherein the feed comprises base metal, Rh in a 3+ oxidation state, Ir in a 4+ oxidation state, and Ru in the form of RuNO, and wherein the concentration of the eluent is reduced prior to elution of the Ir from the column. 14 . A method according to any one of claims 1 to 9 , wherein the feed comprises base metal, one or more of Rh, Ir, and Ru in a 3+ oxidation state, palladium, platinum, and one or more of Fe, Te, Sn, Au, and Os, and wherein the concentration of the eluent is reduced after elution of the base metal, Rh, Ir, and Ru and prior to elution of the palladium and platinum, and wherein the concentration of eluent is further reduced after elution of the palladium and platinum and prior to elution of the Fe, Te, Sn, Au, and/or Os. 15 . A method of preparing a chromatography column for use in the method according to any preceding claims, the method comprising: i) suspending a polymeric chromatography support media in a solution in which the polymeric chromatographic support media is in a contracted state; ii) packing the polymeric chromatography support media into a column while the polymeric chromatographic support media is in its contracted state; iii) adjusting the column while the polymeric chromatographic support media is in its contracted state to restrict the polymeric chromatography support media from moving along the column; and iv) flowing a solution in which the polymeric chromatographic support media expands, through the polymeric chromatography support media in the column, whereby the polymeric chromatographic support media is subjected to compression within the column as it expands and is restricted from moving along the column. 16 . A method according to claim 15 , wherein after step ii) and before step iii), a solution in which the polymeric chromatographic support media is in its contracted state is flowed through the polymeric chromatography support media in the column at a flow rate which provides a pressure drop across the polymeric chromatography support media whereby the polymeric chromatography support media is under compression prior to the adjusting step. 17 . A method according to claim 15 or 16 , wherein adjusting the column in step iii) comprises contacting the polymeric chromatographic support media in the column with an inlet frit and locking the inlet frit in place to restrict the polymeric chromatography support media from moving along the column when subjected to a solution which causes expansion of the polymeric chromatographic support media. 18 . A method according to any one of claims 15 to 17 , further comprising after step iv): v) flowing a solution in which the polymeric chromatographic support media contracts through the polymeric chromatography support media in the column; followed by vi) flowing a solution in which the polymeric chromatographic support media expands through the polymeric chromatography support media in the column. 19 . A method according to any one of claims 15 to 18 , wherein the polymeric chromatographic support media comprises a methacrylate. 20 . A method according to any one of claims 15 to 19 , wherein the polymeric chromatographic support media comprises a glycol. 21 . A method according to any one of claims 15 to 20 , wherein the polymeric chromatographic support media is a co-polymer. 22 . A method according to any one of claims 15 to 21 , wherein the solutions used for contracting and expanding the polymeric chromatographic support media comprise a halide salt or an acid halide, the solutions having different concentrations for contracting and expanding the polymeric chromatographic support media. 23 . A method according to claim 22 , wherein the solutions are HCl, increasing the concentration of the HCl causes the polymeric chromatographic support media to expand, and decreasing the concentration of the HCl causes the polymeric chromatographic support media to contract. 24 . A method according to claim 23 , wherein the HCl is provided at a concentration of at least 5 M to expand the polymeric chromatographic support media and at a concentration of no more than 1 M to contract the polymeric chromatographic support media.
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