Evaluation of a glucose sensing antibody using weak affinity chromatography.
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 alpha1,6-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.
Engström
Henrik A
HA
School of Pure and Applied Natural Sciences, University of Kalmar, SE-391 82 Kalmar, Sweden.
Johansson
Reine
R
Koch-Schmidt
Per
P
Gregorius
Klaus
K
Ohlson
Sten
S
Bergström
Maria
M
D000911:Antibodies, Monoclonal / Q000276:immunology*; D002846:Chromatography, Affinity / Q000379:methods*; D004797:Enzyme-Linked Immunosorbent Assay; D005947:Glucose / Q000032:analysis* / Q000276:immunology
D000911:Antibodies, Monoclonal; D005947:Glucose