Process for obtaining antibodies
The present disclosure relates to a method for the manufacture of recombinant antibody molecules comprising culturing a host cell sample transformed with an expression vector encoding a recombinant antibody molecule; adding an extraction buffer to the sample; and subjecting the sample to a heat treatment step; wherein the pH of the sample is detected after addition of the extraction buffer, and optionally adjusted, to ensure that the pH of the sample is 6 to 9 prior to the heat treatment step.
1. A composition comprising a recombinant antibody and an extraction buffer, said composition being obtained from a method comprising culturing a host cell sample transformed with an expression vector encoding a recombinant antibody molecule; separating the host cell sample from medium in which the host cell was cultured; adding an extraction buffer to the separated host cell sample; detecting the pH of the extraction buffer containing the separated host sample after the extraction buffer is added and adjusting the pH of the extraction buffer containing the separated host sample to a pH of 6-9; and subjecting the sample to a heat treatment step, wherein:
a) the recombinant antibody is a VH, VL, VHH, Fab, modified Fab, altered hinge Fab, Fab′, F(ab′) 2 , Fv fragment; a light chain or heavy chain monomer or dimer; a single chain antibody or a dual specificity antibody; and
b) the recombinant antibody is produced in bacterial cells, yeast cells, Chinese Hamster Ovary (CHO) cells, myeloma cell lines, or hybridoma cell lines.
2. The composition according to claim 1 , wherein said heat treatment step is performed within the range of 30° C. to 70° C.
3. The composition according to claim 1 , wherein the antibody specifically binds to an antigen selected from TNF-α, VLA-4, E-selectin, P selectin, L-selectin, CD2, CD3, CD4, CD5, CD7, CD8, CD11a, CD11b, CD18, CD19, CD20, CD23, CD25, CD33, CD38, CD40, CD40L, CD45, CDW52, CD69, CD134 (OX40), ICOS, BCMP7, CD137, CD27L, CDCP1, CSF1, CSF1-Receptor, DPCR1, DPCR1, dudulin2, FLJ20584, FLJ40787, HEK2, KIAA0634, KIAA0659, KIAA1246, KIAA1455, LTBP2, LTK, MAL2, MRP2, nectin-like2, NKCC1, PTK7, RAIG1, TCAM1, SC6, BCMP101, BCMP84, BCMP11, DTD, carcinoembryonic antigen (CEA), human milk fat globulin (HMFG1 and 2), MHC Class I antigens, MHC Class II antigens, KDR or VEGF.
4. The composition according to claim 1 , wherein the antibody molecule is selected from a VH, VL, VHH, Fab, modified Fab, altered hinge Fab, Fab′, F(ab′) 2 , Fv fragment; a light chain or heavy chain monomer or dimer; a single chain antibody or a dual specificity antibody.
5. The composition according to claim 1 , wherein the pH is adjusted, to ensure that the pH of the sample is 6 to 9 prior to the heat treatment step.
6. The composition according to claim 1 , wherein said antibody binds to TNF-α.
7. The composition according to claim 6 , wherein the antibody is selected from a VH, VL, VHH, Fab, modified Fab, altered hinge Fab, Fab′, F(ab′) 2 , Fv fragment; a light chain or heavy chain monomer or dimer; a single chain antibody or a dual specificity antibody.
8. The composition according to claim 7 , wherein said antibody is a Fab′.
9. The composition according to claim 2 , wherein said heat treatment step is performed within the range of 40° C. to 65° C.
10. The composition according to claim 2 , wherein said heat treatment step is performed within the range of 45° C. to 60° C.
11. The composition according to claim 1 , wherein said heat treatment step is performed for a period of between 6 and 16 hours.
12. The composition according to claim 1 , wherein said heat treatment step is performed for a period of between 4 and 18 hours.
13. The composition according to claim 1 , wherein the extraction buffer is a Tris(hydroxymethyl)aminomethane/Ethylenedinitrilotetraacetic acid disodium salt dehydrate (Tris/EDTA) buffer.