Development of retinol-binding protein 4 immunocolloidal gold fast test strip using high-sensitivity monoclonal antibodies generated by DNA immunization

摘要

DNA immunization is an efficient method for high-affinity monoclonal antibody generation. Here, we describe the gen-eration of several high-quality monoclonal antibodies (mAbs) against retinol-binding protein 4 (RBP4), an important marker for kidney abnormality and dysfunction, with a combination method of DNA priming and protein boost.The mAbs generated could bind to RBP4 with high sensi-tivity and using these mAbs, an immunocolloidal gold fast test strip was constructed. The strip can give a result in <5 min and is very sensitive with a detection limit of about 1 ng/ml. A small-scale cfinical test revealed that the result of this strip was well in accordance with that of an enzymelabeled immunosorbent assay kit currently available on the market. Consequently, it could be useful for more con-venient and faster RBP4 determination in the clinic.