Phage-Mediated Competitive Chemiluminescent Immunoassay for Detecting Cry1Ab Toxin by Using an Anti-Idiotypic Camel Nanobody

Overview

Journal J Agric Food Chem

Specialties Chemistry
Nutritional Sciences

Date 2018 Jan 3

PMID 29293334

Citations 6

Authors

YuLou Qiu

Pan Li

Sa Dong

Xiaoshuai Zhang

Qianru Yang

Yulong Wang

Jing Ge

Bruce D Hammock

Cunzheng Zhang

Xianjin Liu

Affiliations

Soon will be listed here.

Abstract

Cry toxins have been widely used in genetically modified organisms for pest control, raising public concern regarding their effects on the natural environment and food safety. In this work, a phage-mediated competitive chemiluminescent immunoassay (c-CLIA) was developed for determination of Cry1Ab toxin using anti-idiotypic camel nanobodies. By extracting RNA from camels' peripheral blood lymphocytes, a naive phage-displayed nanobody library was established. Using anti-Cry1Ab toxin monoclonal antibodies (mAbs) against the library for anti-idiotypic antibody screening, four anti-idiotypic nanobodies were selected and confirmed to be specific for anti-Cry1Ab mAb binding. Thereafter, a c-CLIA was developed for detection of Cry1Ab toxin based on anti-idiotypic camel nanobodies and employed for sample testing. The results revealed a half-inhibition concentration of developed assay to be 42.68 ± 2.54 ng/mL, in the linear range of 10.49-307.1 ng/mL. The established method is highly specific for Cry1Ab recognition, with negligible cross-reactivity for other Cry toxins. For spiked cereal samples, the recoveries of Cry1Ab toxin ranged from 77.4% to 127%, with coefficient of variation of less than 9%. This study demonstrated that the competitive format based on phage-displayed anti-idiotypic nanobodies can provide an alternative strategy for Cry toxin detection.

Citing Articles

One-Pot Synthesis of HRP&SA/ZIF-8 Nanocomposite and Its Application in the Detection of Insecticidal Crystalline Protein Cry1Ab.

Ye R, Chen H, Li H Nanomaterials (Basel). 2022; 12(15).

PMID: 35957109 PMC: 9370751. DOI: 10.3390/nano12152679.

Recombinant antibodies and their use for food immunoanalysis.

Peltomaa R, Barderas R, Benito-Pena E, Moreno-Bondi M Anal Bioanal Chem. 2021; 414(1):193-217.

PMID: 34417836 PMC: 8380008. DOI: 10.1007/s00216-021-03619-7.

Ultrasensitive and rapid detection of ochratoxin A in agro-products by a nanobody-mediated FRET-based immunosensor.

Tang Z, Liu X, Su B, Chen Q, Cao H, Yun Y J Hazard Mater. 2019; 387:121678.

PMID: 31753666 PMC: 7990105. DOI: 10.1016/j.jhazmat.2019.121678.

Hapten Synthesis, Antibody Development, and a Highly Sensitive Indirect Competitive Chemiluminescent Enzyme Immunoassay for Detection of Dicamba.

Huo J, Barnych B, Li Z, Wan D, Li D, Vasylieva N J Agric Food Chem. 2019; 67(20):5711-5719.

PMID: 31042038 PMC: 6873229. DOI: 10.1021/acs.jafc.8b07134.

Development of a Highly Sensitive Direct Competitive Fluorescence Enzyme Immunoassay Based on a Nanobody-Alkaline Phosphatase Fusion Protein for Detection of 3-Phenoxybenzoic Acid in Urine.

Huo J, Li Z, Wan D, Li D, Qi M, Barnych B J Agric Food Chem. 2018; 66(43):11284-11290.

PMID: 30293433 PMC: 6442738. DOI: 10.1021/acs.jafc.8b04521.

References

De Genst E, Silence K, Decanniere K, Conrath K, Loris R, Kinne J . Molecular basis for the preferential cleft recognition by dromedary heavy-chain antibodies. Proc Natl Acad Sci U S A. 2006; 103(12):4586-91. PMC: 1450215. DOI: 10.1073/pnas.0505379103. View

Shu M, Xu Y, Wang D, Liu X, Li Y, He Q . Anti-idiotypic nanobody: A strategy for development of sensitive and green immunoassay for Fumonisin B₁. Talanta. 2015; 143:388-393. DOI: 10.1016/j.talanta.2015.05.010. View

Soberon M, Gill S, Bravo A . Signaling versus punching hole: How do Bacillus thuringiensis toxins kill insect midgut cells?. Cell Mol Life Sci. 2009; 66(8):1337-49. PMC: 11131463. DOI: 10.1007/s00018-008-8330-9. View

Xu C, Zhang C, Zhong J, Hu H, Luo S, Liu X . Construction of an Immunized Rabbit Phage Display Library for Selecting High Activity against Bacillus thuringiensis Cry1F Toxin Single-Chain Antibodies. J Agric Food Chem. 2017; 65(29):6016-6022. DOI: 10.1021/acs.jafc.7b01985. View

Gruber H, Paul V, Meyer H, Muller M . Determination of insecticidal Cry1Ab protein in soil collected in the final growing seasons of a nine-year field trial of Bt-maize MON810. Transgenic Res. 2011; 21(1):77-88. DOI: 10.1007/s11248-011-9509-7. View

Wang Y, Zhang X, Zhang C, Liu Y, Liu X . Isolation of single chain variable fragment (scFv) specific for Cry1C toxin from human single fold scFv libraries. Toxicon. 2012; 60(7):1290-7. DOI: 10.1016/j.toxicon.2012.08.014. View

Qiu Y, He Q, Xu Y, Bhunia A, Tu Z, Chen B . Deoxynivalenol-mimic nanobody isolated from a naïve phage display nanobody library and its application in immunoassay. Anal Chim Acta. 2015; 887:201-208. DOI: 10.1016/j.aca.2015.06.033. View

Liu Y, Jiang D, Lu X, Wang W, Xu Y, He Q . Phage-Mediated Immuno-PCR for Ultrasensitive Detection of Cry1Ac Protein Based on Nanobody. J Agric Food Chem. 2016; 64(41):7882-7889. DOI: 10.1021/acs.jafc.6b02978. View

Guertler P, Paul V, Albrecht C, Meyer H . Sensitive and highly specific quantitative real-time PCR and ELISA for recording a potential transfer of novel DNA and Cry1Ab protein from feed into bovine milk. Anal Bioanal Chem. 2009; 393(6-7):1629-38. DOI: 10.1007/s00216-009-2667-2. View

10.

Li J, Xu Q, Wei X, Hao Z . Electrogenerated chemiluminescence immunosensor for Bacillus thuringiensis Cry1Ac based on Fe3O4@Au nanoparticles. J Agric Food Chem. 2013; 61(7):1435-40. DOI: 10.1021/jf303774x. View

11.

Bravo A, Gill S, Soberon M . Mode of action of Bacillus thuringiensis Cry and Cyt toxins and their potential for insect control. Toxicon. 2007; 49(4):423-35. PMC: 1857359. DOI: 10.1016/j.toxicon.2006.11.022. View

12.

Gao H, Wen L, Wu Y, Fu Z, Wu G . An ultrasensitive label-free electrochemiluminescent immunosensor for measuring Cry1Ab level and genetically modified crops content. Biosens Bioelectron. 2017; 97:122-127. DOI: 10.1016/j.bios.2017.04.033. View

13.

Jerne N, Roland J, Cazenave P . Recurrent idiotopes and internal images. EMBO J. 1982; 1(2):243-7. PMC: 553027. DOI: 10.1002/j.1460-2075.1982.tb01154.x. View

14.

Paul V, Steinke K, Meyer H . Development and validation of a sensitive enzyme immunoassay for surveillance of Cry1Ab toxin in bovine blood plasma of cows fed Bt-maize (MON810). Anal Chim Acta. 2007; 607(1):106-13. DOI: 10.1016/j.aca.2007.11.022. View

15.

HAMERS-CASTERMAN C, Atarhouch T, Muyldermans S, Robinson G, Hamers C, Songa E . Naturally occurring antibodies devoid of light chains. Nature. 1993; 363(6428):446-8. DOI: 10.1038/363446a0. View

16.

Shu M, Xu Y, Liu X, Li Y, He Q, Tu Z . Anti-idiotypic nanobody-alkaline phosphatase fusion proteins: Development of a one-step competitive enzyme immunoassay for fumonisin B1 detection in cereal. Anal Chim Acta. 2016; 924:53-59. DOI: 10.1016/j.aca.2016.03.053. View

17.

Dong S, Zhang X, Liu Y, Zhang C, Xie Y, Zhong J . Establishment of a sandwich enzyme-linked immunosorbent assay for specific detection of Bacillus thuringiensis (Bt) Cry1Ab toxin utilizing a monoclonal antibody produced with a novel hapten designed with molecular model. Anal Bioanal Chem. 2017; 409(8):1985-1994. DOI: 10.1007/s00216-016-0146-0. View

18.

Qiu Y, He Q, Xu Y, Wang W, Liu Y . Modification of a deoxynivalenol-antigen-mimicking nanobody to improve immunoassay sensitivity by site-saturation mutagenesis. Anal Bioanal Chem. 2015; 408(3):895-903. DOI: 10.1007/s00216-015-9181-5. View

19.

Roda A, Mirasoli M, Guardigli M, Michelini E, Simoni P, Magliulo M . Development and validation of a sensitive and fast chemiluminescent enzyme immunoassay for the detection of genetically modified maize. Anal Bioanal Chem. 2006; 384(6):1269-75. DOI: 10.1007/s00216-006-0308-6. View

20.

Xu C, Zhang X, Liu X, Liu Y, Hu X, Zhong J . Selection and application of broad-specificity human domain antibody for simultaneous detection of Bt Cry toxins. Anal Biochem. 2016; 512:70-77. DOI: 10.1016/j.ab.2016.08.012. View