Antibody labeling with Remazol Brilliant Violet 5R, a vinylsulphonic reactive dye

FERRARI, ALEJANDRO; FRIEDRICH, ADRIÁN D.; WOLMAN, FEDERICO; LEONI, JULIANA

JOURNAL OF IMMUNOASSAY AND IMMUNOCHEMISTRY

TAYLOR & FRANCIS INC

Lugar: Londres; Año: 2012

1532-1819

ABSTRACTColloidal gold is the first choice for labeling antibodies to be used in Point OfCare Testing. However, there are some recent reports on a family of textile dyes ?named ´reactive dyes´? being suitable for protein labeling. In the present work, proteinlabeling conditions were optimized for Remazol Brilliant Violet 5R, and the sensitivityof the labeled antibodies was assessed and compared with that of colloidal-goldlabeled antibodies. Also, the accelerated stability was explored. Optimal conditionswere pH 10.95, dye:Ab molar ratio of 264 and an incubation time of 132 minutes.Labeled antibodies were stable, and could be successfully used in a slot blot assay,detecting as low as 400 ng/ml. Therefore, the present work demonstrates thatvinylsulphonic reactive dyes can be successfully used to label antibodies, and areexcellent candidates for the construction of a new generation of Point of Care Testingkits.