Enzyme-linked immunosorbent assay for the quantitative/qualitative analysis of plant secondary metabolites

Abstract

Immunoassays are antibody-based analytical methods for quantitative/qualitative analysis. Since the principle of immunoassays is based on specific antigen–antibody reaction, the assays have been utilized worldwide for diagnosis, pharmacokinetic studies by drug monitoring, and the quality control of commercially available products. Berson and Yalow were the first to develop an immunoassay, known as radioimmunoassay (RIA), for detecting endogenous plasma insulin [1], a development for which Yalow was awarded the Nobel Prize in Physiology or Medicine in 1977. Even today, after half a century, immunoassays are widely utilized with some modifications from the originally proposed system, e.g., radioisotopes have been replaced with enzymes because of safety concerns regarding the use of radioactivity, which is referred to as enzyme immunoassay/enzyme-linked immunosorbent assay (ELISA). In addition, progress has been made in ELISA with the recent advances in recombinant DNA technology, leading to increase in the range of antibodies, probes, and even systems. This review article describes ELISA and its applications for the detection of plant secondary metabolites.

Preincubation format for a sensitive immunochromatographic assay for monocrotaline, a toxic pyrrolizidine alkaloid

“…Rapidity, simplicity, and cost‐performance are important factors for analytical methods, as well as sensitivity and specificity, when the methods are used in practical applications. PI‐ICA is based on an antibody–antigen reaction that enables the development of sensitive and specific analyses . When considering a rapid analysis that can be performed in 20 min, the PI‐ICA technique developed in this study has significant potential as a detection tool for trace MCT contamination in food and herbal materials because it is simple, inexpensive, and most importantly, highly sensitive and requires no special devices.…”

Preparation of a highly specific single chain variable fragment antibody targeting miroestrol and its application in quality control of Pueraria candollei by enzyme‐linked immunosorbent assay

“…Immunoassays are alternative high throughput analytical methods for quantification of plant bioactive compounds which have gained increasing interest during the past decades. Several antibody‐based immunoassays to determine plant secondary metabolites have been reported . Owing to the specific molecular recognition of antigens and antibodies, immunoassays are known for their high sensitivity and specificity which are suitable for determination of phytochemicals especially the minor bioactive compounds.…”

“…Since the emergence of DNA recombinant technology, various forms of antibodies can be produced by protein expression in bacteria and other host organisms. Recombinant antibodies have advantages over conventional PAbs and MAbs for their simple and rapid process of production and the ability to modify their properties via DNA recombinant technology . Single‐chain variable fragment (scFv) antibody is an engineered antibody consisting of variable regions of heavy (VH) and light (VL) chains linked together with a flexible peptide linker (Figure ).…”

“…Several antibody-based immunoassays to determine plant secondary metabolites have been reported. 16,17 Owing to the specific molecular recognition of antigens and antibodies, immunoassays are known for their high sensitivity and specificity which are suitable for determination of phytochemicals especially the minor bioactive compounds. Previously, our group successfully prepared polyclonal antibodies (PAbs) targeting miroestrol and developed an enzyme-linked immunosorbent assay (ELISA) 18 and an enhanced chemiluminescence (ECL) ELISA.…”

“…Nevertheless, characteristics of PAbs obtained from different batches tend to be varied. 17 In this regard, we also prepared monoclonal antibodies (MAbs) targeting miroestrol by fusion of myeloma cells and splenocytes of mice immunised with miroestrol immunogen conjugate to yield hybridoma cells secreting anti-miroestrol MAbs and developed a MAb-based ELISA to determine miroestrol in plant samples. 21 Since the emergence of DNA recombinant technology, various forms of antibodies can be produced by protein expression in bacteria and other host organisms.…”