separation of biotin labelled IgG from unlabelled IgG
Carlos.Duarte at CIGB.EDU.CU
Tue Feb 2 20:01:38 EST 1999
For most applications of biotynilated IgG you dont need to separated unlabel antibodies since the reaction is quite efficient
and you use an excess of biotin in the coupling reaction. Anyway biotinylated Abs will certainly binds to an
affinity column with avidin or streptavidine but you will find very difficult to elute the molecule due to the very high Ka between
biotine and avidine (10-15M). Hope it helps
Carlos A. Duarte, pHD
Head, AIDS Department, Vaccine Division
Center for Genetic Engineering and Biotechnology
Ave 31 entre 158 y 190, Cubanacán, Playa
Apdo 6162, La Habana 10600
Fax 53-7-214764, 218070. 336008
Phone 53-7-218164, 218466, 218008
email carlos.duarte at cigb.edu.cu
From: Dan Pashniak [SMTP:dpashnia at cangene.com]
Sent: Tuesday, February 02, 1999 12:32 PM
To: methods at net.bio.net
Subject: separation of biotin labelled IgG from unlabelled IgG
Hi there . . .
I am looking for a method of separating hapten-labeled IgG, specifically, biotin labeled,
from unlabeled IgG. I am considering using an affinity column bound with anti-biotin.
It's simple to remove the unbound biotin from the reaction matrix . . . just use a good old
PD-10 (G25) column (Pharmacia). But, what about the unlabeled IgG. Size exclusion certainly won't work; biotin is only ~250 Da.
Would and affinity column work?
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