Patent ID: 11919924
Assignee: FRESENIUS KABI DEUTSCHLAND GMBH
Field: Chemical engineering (Chemistry)
Classification: CPC C  A  B | IPC A  B  C

Claim 11:
12. A method of producing an antibody having (i) a light chain sequence comprising SEQ ID NO:1 and a heavy chain sequence comprising SEQ ID NO:2 comprising the steps of:
(a) expressing the antibody in CHO cells and secreting the antibody into a cell culture media to obtain a sample;
(b) contacting the antibody with a hydroxyapatite resin under conditions such that about 25 mg to about 40 mg of the antibody per liter of resin binds to the resin and at least a portion of the impurities remain unbound to the resin, wherein the conditions such that the antibody binds to the resin comprise the use of a loading buffer, wherein the loading buffer: (i) has a pH of about pH 7.5; and (ii) comprises about 5 mM NaPO4;
(c) washing the resin with a wash buffer under conditions such that at least a portion of the impurities are separated from the antibody; and
(d) eluting the antibody from the resin under conditions such that the antibody becomes unbound from the resin to produce an eluate, wherein the eluate contains a lower level of impurities than the sample of step (a), wherein the conditions such that the antibody becomes unbound from the resin comprise the use of an elution buffer, wherein the elution buffer: (i) has a pH of about pH 7.5; (ii) comprises about 10 mM NaPO4; and (iii) comprises about 350 mM NaCl,
wherein about means within 3% of the given value.