AFFINITY MEDIUM USING FIXED WHOLE CELLS
Separation media comprising a support and crosslinked whole cells fixed to the support. The separation media are useful in affinity columns for the separation of the antibodies from a solution, particularly for the separation of polyclonal antibodies that have been raised against the same type of whole cells. In an embodiment, the whole cells are whole bacterial cells.
1 . A support comprising whole cells that are crosslinked to the support.
2 . The support of claim 1 , wherein the whole cells are crosslinked with methylene bridges between surface proteins of the whole cells.
3 . The support of claim 1 , wherein the whole cells are fixed with formalin.
4 . The support of claim 1 , wherein the whole cells are bacterial cells.
5 . The support of claim 4 , wherein the bacterial cells are selected from the group consisting of E. coli, Listeria, Clostridium, Mycobacterium, Shigella, Borrelia, Campylobacter, Bacillus, Salmonella, Staphylococcus, Enterococcus, Pneumococcus, Streptococcus , and a combination thereof.
6 . The support of claim 1 , wherein the support comprises polymer beads or a polymer matrix.
7 . The support of claim 1 , wherein the support comprises silica beads or a silica matrix.
8 . The support of claim 1 , wherein the support is a well plate.
9 . An affinity column comprising a support and crosslinked whole cells that are fixed to the support.
10 . The affinity column of claim 9 , wherein the whole cells are crosslinked with methylene bridges between surface proteins of the whole cells.
11 . The affinity column of claim 9 , wherein the whole cells are fixed with formalin.
12 . The affinity column of claim 9 , wherein the whole cells are bacterial cells.
13 . The affinity column of claim 12 , wherein the bacterial cells are selected from the group consisting of E. coli, Listeria, Clostridium, Mycobacterium, Shigella, Borrelia, Campylobacter, Bacillus, Salmonella, Staphylococcus, Enterococcus, Pneumococcus, Streptococcus , and a combination thereof.
14 . The affinity column of claim 9 , wherein the support comprises polymer beads or a polymer matrix.
15 . The affinity column of claim 9 , wherein the support comprises silica beads or a silica matrix.
16 . A method for purifying antibodies from a solution, the method comprising:
contacting a solution comprising antibodies to a support comprising crosslinked whole cells that are fixed to the support under conditions such that at least one antibody binds the whole cells;
separating the bound antibodies from unbound antibodies; and
eluting the bound antibodies from the support.
17 . The method of claim 16 , wherein the whole cells are crosslinked with methylene bridges between surface proteins of the whole cells.
18 . The method of claim 16 , wherein the whole cells are fixed with formalin.
19 . The method of claim 16 , wherein the whole cells are bacterial cells.
20 . The method of claim 19 , wherein the bacterial cells are selected from the group consisting of E. coli, Listeria, Clostridium, Mycobacterium, Shigella, Borrelia, Campylobacter, Bacillus, Salmonella, Staphylococcus, Enterococcus, Pneumococcus, Streptococcus , and a combination thereof.
21 . The method of claim 16 , wherein the solution comprises monoclonal or polyclonal antibodies.
22 . The method of claim 21 , wherein the antibodies are from a mouse, a rabbit, or a goat.
23 . The method of claim 16 , wherein the molar ratio of antibodies to whole cell antigens is at least about 1:2.
24 . The method of claim 16 , further comprising:
contacting serum comprising the antibodies with a protein affinity column to bind the antibodies to the protein;
washing the protein affinity column with the antibodies bound thereto; and
eluting the antibodies from the protein affinity column, thereby creating a solution comprising antibodies.
25 . The method of claim 24 , wherein the protein is protein G.
26 . The method of claim 16 , wherein eluting the antibodies from the separation medium comprises contacting the antibodies bound to the separation medium with an elution buffer.