Development of an Enzyme Linked Immunosorbent Assay for Direct Determination of Anticancer Drug Vitamin K3 in Serum

Abstract

Synthetic vitamin K3 (VK3, 2-methyl-1,4-naphthoquinone, or menadione) has been shown to have anticancer activity in various human cancer cells. We developed an enzyme-linked immunosorbant assay (ELISA) for determination of VK3 in fetal calf serum (FCS) without any pretreatment. The monoclonal antibody against VK3, which was secreted from an established hybridoma cell line (3A3) has been proven to have highly specific binding to VK3 in cross-reactivity analysis. The full measuring range of the assay extends from 0.39 to 50 µg/ml of VK3. Based on validation analysis, this immunological analysis is a precise, accurate, and sensitive method for determination of VK3. This simple immunological method would deserve to be applicable clinically.

Synthesis and Characterization of Antibody-Protected Bimetallic Nanoclusters with Catalytic Properties

“…Immunoassays are analytical techniques in which the selectivity and sensitivity are provided by the specific interaction between an antibody and its target analyte . Enzyme-linked immunosorbent assay (ELISA), taking advantage of enzymes attached to the antibodies such as horseradish peroxidase (HRP) or alkaline phosphatase (ALP) for signal amplification, is the most frequently employed method in clinical diagnostics. − Natural enzymes as markers have several disadvantages, such as, high susceptibility to environmental variations, easy denaturation and digestion, and costly and time-consuming preparation and purification . Enzymes and antibodies are incubated with cross-linking reagents and then the conjugate is purified.…”

Comparison of chemiluminescence enzyme immunoassay based on magnetic microparticles with traditional colorimetric ELISA for the detection of serum α-fetoprotein

“…The most common detection method of EIA is enzyme-linked immunosorbent assay (ELISA). ELISA was first reported by Engvall and Perlmann in 1971 [3] and has been widely used as a diagnostic tool in clinic [4] , [5] , plant pathology [6] and food industry [7] . However, the need to detect increasingly smaller amounts of target molecules has led to the emergence of more sensitive indicators, such as fluorescence and chemiluminescence.…”

Analytical techniques for the determination of biologically active quinones in biological and environmental samples

“…The immunosensor could determine DXR in human serum without interference from biological components. An indirect competitive enzyme linked immunosorbent assay (ELISA) was developed for the specific determination of menadione (2-methyl-1, 4-naphthoquinone) [82]. The monoclonal antibody used in the assay was prepared by ovalbumin modified with plumbagin (2-hydroxy-1, 4naphthoquinone) as a hapten protein.…”