Molecular engineering of biotin-glutamic acid decarboxylase 65 fusion protein (Biotin-GAD65) for non-radioactive GAD65 antibody assay.
We constructed biotinylated fusion proteins that linked to three detection tags to GAD65 at the N-terminus, and expressed them in an E. coli expression system. The Biotin14-GAD65 protein exhibited the strongest binding to both the GAD65 antibody and the streptavidin among the three constructs. We describe the optimal conditions using a Biotin14-GAD65-based immunoassay for the detection of GAD65 antibody.
Luo
Dong
D
Department of Medicine, University of Washington, Box 357710, HSB K-165, 1959 Pacific Street N.E., Seattle, WA 98195-7710, USA.
Rogers
Chad N
CN
Hampe
Christiane S
CS
Lernmark
Ake
A
D000906:Antibodies / Q000032:analysis* / Q000276:immunology; D000936:Antigen-Antibody Complex / Q000032:analysis; D015374:Biosensing Techniques / Q000379:methods*; D001710:Biotin / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005968:Glutamate Decarboxylase / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D007527:Isoenzymes / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D011485:Protein Binding; D015202:Protein Engineering / Q000379:methods*; D011863:Radioimmunoassay; D011993:Recombinant Fusion Proteins; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D000906:Antibodies; D000936:Antigen-Antibody Complex; D007527:Isoenzymes; D011993:Recombinant Fusion Proteins; D001710:Biotin; D005968:Glutamate Decarboxylase; C401141:glutamate decarboxylase 2