Immunoassay based on a polyclonal antibody for sex steroid hormones produced by a heterogeneous hapten-conjugated immunogen: Estimation of its potentiality and antibody characteristics

Watanabe, Eiji; Kubo, Hiroaki; Kanzaki, Yukiko; Nakazawa, Hiroyuki

doi:10.1016/j.aca.2009.10.050

Cited by 8 publications

(3 citation statements)

References 9 publications

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“…Immunodetection based on the specific interaction between antigens and antibodies has been widely used in pharmaceutical diagnosis, medical therapy, and fundamental researches. 1,2 Enzyme-linked immunosorbent assay (ELISA), the most popular immunoassay method, provides a rapid and efficient screening for target molecules such as drugs, 3,4 herbal compounds, 5 pesticides, 6 herbicides, 7 antibiotics, 8 hormones, 9 and marker proteins of diseases. [10][11][12] The key point to develop immunoassay for the detection of small compounds is to obtain their antibodies with high specificity and affinity.…”

Section: Introductionmentioning

confidence: 99%

Engineering lysine‐rich caleosins as carrier proteins to render biotin as a hapten on artificial oil bodies for antibody production

Liu

Chyan

et al. 2011

Biotechnology Progress

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It has been demonstrated that caleosin alone is sufficient to stabilize artificial oil bodies. A series of recombinant caleosins, mutated with 3, 5, 8, 11, 13, 15, and 17 extra Lys residues and over-expressed in Escherichia coli, were used as carrier proteins to render biotin as a hapten on the surface of artificial oil bodies for antibody production. Biotinylation levels of the recombinant caleosins were step-wisely elevated as the number of extra Lys residues increased, and the biotinylated Lys residues were identified by mass spectrometric analysis. Polyclonal antibodies against biotin were successfully generated in rats injected with artificial oil bodies constituted with each of the biotinylated caleosins. Moreover, those generated via the biotinylated caleosins with eight or more extra Lys residues no longer recognized caleosin. It appears that engineered Lys-rich caleosins are suitable carrier proteins for the production of antibodies against small molecules. (C) 2011 American Institute of Chemical Engineers Biotechnol. Prog., 201

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Section: Introductionmentioning

confidence: 99%

Engineering lysine‐rich caleosins as carrier proteins to render biotin as a hapten on artificial oil bodies for antibody production

Liu

Chyan

et al. 2011

Biotechnology Progress

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“…It would produce polyclonal antisera that can recognize more than one compound at a time. Watanabe et al [28] designed the hapten molecule of sex steroid hormones in which the linker group is attached to the D-ring; it would be the best approach to produce desirable antibodies that can distinguish both compounds (17β-estradiol and testosterone).…”

Section: Hapten: Design and Synthesismentioning

confidence: 99%

Antibody production and application for immunoassay development of environmental hormones: a review

Tian

Wang

Lei

et al. 2018

Chem. Biol. Technol. Agric.

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Environmental hormones, also called environmental endocrine-disrupting chemicals (EDCs), are produced by various human activities and environment pollution, and they are composed of industrial chemicals and environmental pollutants. They play a harmful role in breaking the balance of normal physiological metabolism, causing serious diseases and disrupting reproductive development in wildlife and human beings by inhibiting or imitating the action of normal gonadal hormones. Strict controls of maximum residue levels to some environmental hormones in foodstuff have been set up by many countries. Traditional detection methods, due to their high accuracy and maturity, are used extensively, such as HPLC, HPLC-MS, and GC-MS. However, these methods are time consuming and require extraordinary skills of operators, and sometimes they cannot meet the requirements of field testing. Immunoassay has the advantages of high specificity, sensitivity, simplicity, convenience, and the ability to achieve high throughput, playing an irreplaceable role in the field of rapid detection. This review focuses on the antibody production and the development of immunoassays to detect environmental hormones in food and environment. Therefore, estrogen, progesterone, and testosterone are selected as examples to introduce a series of procedures for the detection of environmental hormones including antigen synthesis, antibody production, and antibody-based detection methods.

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“…Actually, there were reports showing that the antibody obtained from at 3-position were more sensitive than that obtained from 17-position of E 2 . Watanabe, Kubo, Kanzaki, and Nakazawa (2010) previously prepared β-estradiol 17-hemisuccuinate as a hapten for producing polyclonal antibodies with an IC 50 values of 6.9 ng/mL, and Yang et al (2015) synthesized E 2 -CME as a hapten for conjugating carrier proteins, obtaining a monoclonal antibody specific for E 2 with an IC 50 value of 0.18 ng/mL using a lateral-flow colloidal gold immunoassay strip. Thus, we synthesized the E 2 -CME used as hapten to produce antibodies to E 2 in this study.…”

Section: Preparation Of Haptens and Conjugatesmentioning

confidence: 99%

Production of antibodies and development of an enzyme-linked immunosorbent assay for 17β-estradiol in milk

Bai

Liu

et al. 2017

Food and Agricultural Immunology

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The residue of 17β-estradiol (E 2) in milk could potentially lead to the occurrence of various reproductive diseases; therefore, a rapid and sensitive method for monitoring E 2 residues in milk was highly necessary. In this study, we produced new polyclonal and monoclonal antibodies using E 2-3-O-carboxymethyl ether as a hapten and developed an indirect competitive enzyme-linked immunosorbent assay (icELISA) for the detection of E 2 in milk. The results showed that the sensitivity of polyclonal antibody was higher than that of monoclonal antibody, providing a half maximum inhibition concentration (IC 50) against E 2 of 0.17 ng/mL, high cross-reactivity (CR) to E 2 benzoate (150%) and oestriol (18.02%), and negligible CR with other oestrogen compounds. Under optimized conditions, the developed icELISA based on the polyclonal antibody had a limit of detection values of 0.093 μg/L, which was enough sensitive to detect E 2 in milk. In spiked samples (0.5, 1, and 2 μg/L), the recoveries ranged from 83.12% to 94.58% with coefficients of variation <12.8%. These results indicated that the icELISA method we developed was suitable for screening of E 2 residue in milk.

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Immunoassay based on a polyclonal antibody for sex steroid hormones produced by a heterogeneous hapten-conjugated immunogen: Estimation of its potentiality and antibody characteristics

Cited by 8 publications

References 9 publications

Engineering lysine‐rich caleosins as carrier proteins to render biotin as a hapten on artificial oil bodies for antibody production

Engineering lysine‐rich caleosins as carrier proteins to render biotin as a hapten on artificial oil bodies for antibody production

Antibody production and application for immunoassay development of environmental hormones: a review

Production of antibodies and development of an enzyme-linked immunosorbent assay for 17β-estradiol in milk

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