Publications by authors named "Helen Brooking"
Background: GAD(65)Ab are important markers of risk of development of type 1 DM.
Methods: With the need to improve the disease specificity of GAD(65)Ab measurement in mind, we have analysed the interaction between recombinant human GAD(65) and GAD(65)Ab from different groups of subjects in terms of association and dissociation rate constants and equilibrium constants. In addition, binding of GAD(65)Ab from various groups of subjects to wild-type GAD(65) versus GAD(65) containing a mutation E517P was studied.
Background: A sensitive ELISA for measurement of IA-2 autoantibodies has been developed and assessed. Also, a combination ELISA for detection of both GAD65 autoantibodies and IA-2 autoantibodies is described.
Methods: The IA-2 autoantibody assay is based on the ability of IA-2 autoantibodies to form a bridge between IA-2 intracellular fragment coated onto ELISA plate wells and liquid-phase IA-2 labelled with biotin.
Background: A new ELISA for 65 kDa isoform of glutamic acid decarboxylase autoantibodies (GAD(65) Abs), which depends on GAD(65) Ab acting divalently and forming a bridge between immobilized GAD(65) and liquid-phase GAD(65)-biotin, is described.
Methods: Sera (25 microl) were incubated in GAD(65)-coated ELISA plate wells followed by washing and incubation with GAD(65)-biotin. After a further wash step, GAD(65)-biotin bound was quantitated by addition of streptavidin peroxidase followed by tetramethylbenzidine.