title: Production of rapidly reversible antibody and its performance characterization as binder for continuous glucose monitoring.
abstract: To effectively control diabetes, a method to reliably measure glucose fluctuations in the body over given time periods needs to be developed. Current glucose monitoring systems depend on the substrate decomposition by an enzyme to detect the product; however, the enzyme activity significantly decays over time, which complicates analysis. In this study, we investigated an alternative method of glucose analysis based on antigen-antibody binding, which may be active over an extended period of time. To produce monoclonal antibodies, mice were immunized with molecular weight (M(W)) 10K dextran chemically conjugated with keyhole limpet hemocyanin. Since dextran contains glucose molecules polymerized via a 1,6-linkage, the produced antibodies had a binding selectivity that could discriminate biological glucose compounds with a 1,4-linkage. Three antibody clones with different affinities were screened using the M(W) 1K dextran-bovine serum albumin conjugates as the capture ligand. Among the antibodies tested, the antibody clone Glu 26 had the lowest affinity (K(A) = 3.56 × 10(6) M(-1)) and the most rapid dissociation (k(d) = 1.17 × 10(-2) s(-1)) with the polysaccharide immobilized on the solid surfaces. When glucose was added to the medium, the sensor signal was inversely proportional to the glucose concentration in a range between 10 and 1000 mg dL(-1), which covered the clinical range. Under the optimal conditions, the response time was about 3 min for association and 8 min for dissociation based on a 95% recovery of the final equilibrium.
authors: Program for Bio-Microsystem Technology, Korea University, 204C Specific Research Wing, Biotechnology Building (Green Campus), 1, 5-ka, Anam-dong, Sungbuk-gu, Seoul, 136-701, Korea.
authors: Sung-Ho
authors: SH
authors: Paek
authors: Il-Hoon
authors: IH
authors: Cho
authors: Sung-Min
authors: SM
authors: Seo
authors: Dong-Hyung
authors: DH
authors: Kim
authors: Se-Hwan
authors: SH
authors: Paek
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000915:Antibody Affinity; D000937:Antigen-Antibody Reactions; D015374:Biosensing Techniques / Q000379:methods*; D002417:Cattle; D003911:Dextrans / Q000737:chemistry; D005947:Glucose / Q000032:analysis*; D006433:Hemocyanins / Q000737:chemistry / Q000276:immunology; D007700:Kinetics; D008024:Ligands; D051379:Mice; D012710:Serum Albumin, Bovine / Q000737:chemistry
chemical_list: D000911:Antibodies, Monoclonal; D003911:Dextrans; D008024:Ligands; D012710:Serum Albumin, Bovine; D006433:Hemocyanins; C032808:keyhole-limpet hemocyanin; D005947:Glucose