Breaking strepavidin/biotin bond.

marder at agri.huji.ac.il marder at agri.huji.ac.il
Tue Apr 14 03:03:03 EST 1992


In article <9204121706.AA25167 at genbank.bio.net>, MIBJOEL at UKCC.UKY.EDU (Joel Guthridge) writes:
> Tom Schneider recently asked how one could break the strepavidin/biotin
> interaction to recover the biotin conjugated material.  If I remember
> correctly, one can use iminobiotin instead of biotin.  The advantage
> is that the iminobiotin/strepavidin interaction has a much lower affinity
> than the biotin/strepavidin interaction...  allowing less harsh elution
> methods to be employed.  I think Sigma and Pierce (as well as others I'm
> sure) sell NHS- or hydrazine based iminobiotin for conjugate preparation.

I think that you may be in error here;  the main thing about iminobiotin is
that it has a pK around 10 and only one form binds (strept)avidin, tho' I can't
remember whether it is the protonated or unprotonated form.  In either case,
you have to expose your sample to fairly alkaline conditions for the
binding or unbinding stage - not too bad for most proteins, but perhaps
problematic for nucleic acids.  Iminobiotin and derivitive labelling kits
are listed in the Sigma catalogue and they give the following refs:-
	Heney G et al. (1981) Analytical Biochem. 114,92.
	Orr GA et al. (1981) JBC 256,761.
Another biotin derivative listed in the Sigma catalogue is Diaminobiotin
(Ref. Green NM; 1966, Biochem. J., 101, 774).  Perhaps it works in a
similar way.
--
                                 '      Jonathan B. Marder
Internet: MARDER at AGRI.HUJI.AC.IL :      Department of Agricultural Botany
Bitnet:   MARDER at HUJIAGRI        : /\/  The Hebrew University of Jerusalem
Phone:    (08 or +9728) 481918   :/  \  Faculty of Agriculture
Fax:      (08 or +9728) 467763   /      P.O.Box 12, Rehovot 76100, ISRAEL



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