Adsorbing serum

jd89 Jan_B_DEJARNETTE at UMAIL.UMD.EDU
Sat Oct 14 14:20:59 EST 1995


Dear netters,
     I would like to adsorb polyclonal serum to make my antiserum more
monospecific and would like to know if anyone has had success with the
various methods.  The unwanted reactivity in the antiserum is due to
contaminating proteins in the original antigen prep.  The antiserum has a
very high titer and the number of contaminating proteins in the original
antigen prep is low.  The three most notable methods appear to be;  1)
nitrocellulose coated with contaminating protein prep (non-covalent),  2)
diazotized paper treated with contaminating protein prep (covalent), and 3)
pecipitable acetone powder of the contaminating protein prep.  I would like
to be able to adsorb the unwanted antibodies then elute them off with a low
pH and then reuse the absorbing medium.  I'm personally leaning towards the
nitrocellulose (dipstick) method, but would greatly appreciate comments and
advice.  Please send replies to my email address.  Thanks in advance.

                                         Jan DeJarnette
                                       jd89 at umail.umd.edu



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