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A filtration method for rapid preparation of conjugates for immunoassay

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Title A filtration method for rapid preparation of conjugates for immunoassay
 
Creator Dhar, Tarun K
Dasgupta, Subham
Roy, Dipika
Banerjee, Meghnna
 
Subject Drug Development/Diagnostics & Biotechnology
 
Description Modification of protein and other biopolymers by labeling them with small or macromolecules has become a very powerful research tool in biochemistry, molecular biology, diagnostics, and therapeutics. However, current methodologies available for their preparations are not
straightforward and take several hours of incubation time. In this paper, we describe a new filtration-assisted technique for covalent conjugation between the reactive functional groups of two different molecules (small or macromolecules). Compared to the current method, this
new approach significantly reduces the total reaction time from several hours to just a few minutes. The technique has been used for the preparation of conjugates of a small molecule to a protein such as biotin–BSA conjugate or small molecules to a small molecule such as biotin– tyramine conjugate or protein–protein conjugation such as antibody-horseradish peroxidase conjugate. The procedure consists of filtering the reaction mixture multiple times through
membrane micropores with the help of two syringes, which make the cross filtration process less laborious. The method saves time, allows conjugation of less than 1 mg protein and produces conjugates better than those obtained by the current methods. Although the present technique has been applied on some common conjugation methods, it provides a potentially generalmethod, and may further be expanded for the synthesis of several other macromolecular
conjugates
 
Publisher Elsevier
 
Date 2012
 
Type Article
PeerReviewed
 
Format application/pdf
 
Identifier http://www.eprints.iicb.res.in/2675/1/Journal_of_Immunological_Methods_385_(2012).pdf
Dhar, Tarun K and Dasgupta, Subham and Roy, Dipika and Banerjee, Meghnna (2012) A filtration method for rapid preparation of conjugates for immunoassay. Journal of Immunological Methods, 385. pp. 71-18.
 
Relation http://dx.doi.org/10.1016/j.jim.2012.08.009
http://www.eprints.iicb.res.in/2675/