In article <87c9n8$gh0$1 at nnrp1.deja.com>, <URL:mailto:nan at inflam.lu.se> wrote:
> Hello friends,
>> I am using 5-10% of Fetal bovine serum containing medium to culture my B-
> cell hybridomas. While purifying the culture supernatants using Protein G
> column (Gama bind plus), I am facing the problem of bovine
> immunoglobulins copurifying with my monoclonal antibodies. How to remove
> the bovine immunoglobulins contamination from my purified monoclonal
> antibodies? Is there any company marketing affigel columns with anti-
> bovine immunoglobulins? or is there any alternative methods available?
> Now I have started using either protein A passed FBS or Ultra low bovine
> immunoglobulin containing sera for the culture medium to circumvent this
> problem. Any help in this regard will be useful. Thanks in advance for
> your suggestions.
>> regards,
> K.S.Nandakumar
>One way to achieve this is to preadsorb the FBS on Protein A or Protein G.
Of course you will need to keep the column clean (free of bacteria and
pyrogens) and filter the FBS afterwards.
Many commercial companies pass therapeutic antibodies through an
anti-bovine Ig column as a stage of purification.
Mike <URL:http://www.path.cam.ac.uk/~mrc7/>
--
o/ \\ // || ,_ o M.R. Clark, PhD. Division of Immunology
<\__,\\ // __o || / /\, Cambridge University, Dept. Pathology
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