Evaluation of a glucose sensing antibody using weak affinity chromatography

✍ Scribed by Henrik A. Engström; Reine Johansson; Per Koch-Schmidt; Klaus Gregorius; Sten Ohlson; Maria Bergström

• Publisher: John Wiley and Sons
• Year: 2008
• Tongue: English
• Weight: 113 KB
• Volume: 22
• Category: Article
• ISSN: 0269-3879
• DOI: 10.1002/bmc.924

✦ Synopsis

Abstract

Continuous monitoring of drug levels and endogenous molecules in biological fluids is a developing research area with many applications. One example is the need to improve life for millions of diabetes mellitus patients by continuously monitoring the glucose level. In order to have a dynamic response, the recognition molecule in a continuous sensor should preferentially have a fast dissociation rate and a dissociation constant in the millimolar range. We have evaluated the monoclonal antibody (mAb) 3F1E8‐A2 for its potential to be used in a future glucose sensor application. The mAb was generated from hybridomas by immunizing mice with 10 kDa dextran (an __α__1,6‐glucose polymer) with the aim of obtaining mAbs that can recognize the glucose monomer. The mAb was immobilized to macroporous silica and the interaction with dextran‐derived oligosaccharides was evaluated with weak affinity chromatography (WAC). To measure the low affinities between the mAb 3F1E8‐A2 and different monosaccharides, a competitive weak affinity chromatography approach was employed. It was found that the mAb had a higher specificity for glucose compared with other monosaccharides and the dissociation constant (K~d~) towards glucose was determined as 18.8 ± 2.6 mm. Copyright © 2007 John Wiley & Sons, Ltd.