Full Text:   <2304>

Summary:  <1701>

CLC number: Q74

On-line Access: 2016-04-05

Received: 2015-11-11

Revision Accepted: 2015-12-28

Crosschecked: 2016-03-13

Cited: 5

Clicked: 3500

Citations:  Bibtex RefMan EndNote GB/T7714


Yi-rong Guo


-   Go to

Article info.
Open peer comments

Journal of Zhejiang University SCIENCE B 2016 Vol.17 No.4 P.282-293


Evaluation of a water-soluble adjuvant for the development of monoclonal antibodies against small-molecule compounds

Author(s):  Rui Liu, Ying Liu, Mei-jing Lan, Niusha Taheri, Jing-li Cheng, Yi-rong Guo, Guo-nian Zhu

Affiliation(s):  Institute of Pesticide and Environmental Toxicology, Zhejiang University, Hangzhou 310058, China

Corresponding email(s):   yirongguo@zju.edu.cn

Key Words:  Water-soluble adjuvant, Monoclonal antibody, Hapten-specific antibody, High affinity

Rui Liu, Ying Liu, Mei-jing Lan, Niusha Taheri, Jing-li Cheng, Yi-rong Guo, Guo-nian Zhu. Evaluation of a water-soluble adjuvant for the development of monoclonal antibodies against small-molecule compounds[J]. Journal of Zhejiang University Science B, 2016, 17(4): 282-293.

@article{title="Evaluation of a water-soluble adjuvant for the development of monoclonal antibodies against small-molecule compounds",
author="Rui Liu, Ying Liu, Mei-jing Lan, Niusha Taheri, Jing-li Cheng, Yi-rong Guo, Guo-nian Zhu",
journal="Journal of Zhejiang University Science B",
publisher="Zhejiang University Press & Springer",

%0 Journal Article
%T Evaluation of a water-soluble adjuvant for the development of monoclonal antibodies against small-molecule compounds
%A Rui Liu
%A Ying Liu
%A Mei-jing Lan
%A Niusha Taheri
%A Jing-li Cheng
%A Yi-rong Guo
%A Guo-nian Zhu
%J Journal of Zhejiang University SCIENCE B
%V 17
%N 4
%P 282-293
%@ 1673-1581
%D 2016
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.B1500278

T1 - Evaluation of a water-soluble adjuvant for the development of monoclonal antibodies against small-molecule compounds
A1 - Rui Liu
A1 - Ying Liu
A1 - Mei-jing Lan
A1 - Niusha Taheri
A1 - Jing-li Cheng
A1 - Yi-rong Guo
A1 - Guo-nian Zhu
J0 - Journal of Zhejiang University Science B
VL - 17
IS - 4
SP - 282
EP - 293
%@ 1673-1581
Y1 - 2016
PB - Zhejiang University Press & Springer
ER -
DOI - 10.1631/jzus.B1500278

A water-soluble adjuvant named QuickAntibody (QA) was introduced into the procedure of mouse immunization for the development of hapten-specific monoclonal antibodies (mAbs), using four kinds of pesticides as model compounds. Compared with conventional Freund’s adjuvants, QA treatments offered relatively low but acceptable antiserum titers after three inoculations, gave little adverse effects to the experimental animals, and were preferable in harvesting splenocytes during the steps of cell fusion. Afterwards, hybridomas from the QA group were prepared and screened by both non-competitive and competitive indirect enzyme-linked immunosorbent assays (ELISAs). The efficiency of gaining immune-positive hybridomas was satisfactory, and the resultant mAbs showed sensitivities (half maximal inhibitory concentration (IC50)) of 0.91, 2.46, 3.72, and 6.22 ng/ml to triazophos, parathion, chlorpyrifos, and fenpropathrin, respectively. Additionally, the performance of QA adjuvant was further confirmed by acquiring a high-affinity mAb against okadaic acid (IC50 of 0.36 ng/ml) after three immunizations. These newly developed mAbs showed similar or even better sensitivities compared with previously reported mAbs specific to the corresponding analytes. This study suggested that the easy-to-use adjuvant could be applicable to the efficient generation of highly sensitive mAbs against small compounds.


方法:本研究以4种农药和1种海洋毒素为目标分析物,采用一种新型水溶性佐剂与小分子抗原相互 混合,直接对Balb/c小鼠进行小腿肌肉注射免疫。 经过2~3次免疫后进行腹腔注射常规末次免疫,细胞 融合后筛选出了相应的杂交瘤细胞株,制备并鉴定了针对上述半抗原小分子的特异性单抗。


Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article


[1]Algate, P., Baldridge, J., Mossman, S.P., 2013. Antigens and adjuvants. In: Howard, G.C., Kaser, M.R. (Eds.), Making and Using Antibodies: A Practical Handbook. CRC Press, London, p.22-30.

[2]Alving, C.R., Matyas, G.R., Torres, O., et al., 2014. Adjuvants for vaccines to drugs of abuse and addiction. Vaccine, 32(42):5382-5389.

[3]Chen, X., Liu, L., Kuang, H., et al., 2013. A strip-based immunoassay for rapid determination of fenpropathrin. Anal. Methods, 5(21):6234-6239.

[4]Chen, X., Xu, L., Ma, W., et al., 2014. General immunoassay for pyrethroids based on a monoclonal antibody. Food Agric. Immunol., 25(3):341-349.

[5]Erhard, M., Kellner, J., Kühlmann, R., et al., 1991. Influence of various adjuvants on the synthesis of specific antibodies of chicken, sheep and rabbit following immunization with an hapten. J. Vet. Med. A, 38(1-10):21-27.

[6]Esteve-Turrillas, F.A., Mercader, J.V., Agullo, C., et al., 2014. Design and development of heterologous competitive immunoassays for the determination of boscalid residues. Analyst, 139(14):3636-3644.

[7]Ferber, P.C., Ossent, P., Homberger, F.R., et al., 1999. The generation of monoclonal antibodies in mice: influence of adjuvants on the immune response, fusion efficiency and distress. Lab. Anim., 33(4):334-350.

[8]Fodey, T.L., Delahaut, P., Charlier, C., et al., 2008. Comparison of three adjuvants used to produce polyclonal antibodies to veterinary drugs. Vet. Immunol. Immunopathol., 122(1-2):25-34.

[9]George, S.E., Elliott, C.T., McLaughlin, D.P., et al., 2012. An investigation into the potential use of nanoparticles as adjuvants for the production of polyclonal antibodies to low molecular weight compounds. Vet. Immunol. Immunopathol., 149(1-2):46-53.

[10]Gui, W.J., Jin, R.Y., Chen, Z.L., et al., 2006. Hapten synthesis for enzyme-linked immunoassay of the insecticide triazophos. Anal. Biochem., 357(1):9-14.

[11]Guo, Y., Sanders, M., Galvita, A., et al., 2014. Heterologous screening of hybridomas for the development of broad-specific monoclonal antibodies against deoxynivalenol and its analogues. World Mycotoxin J., 7(3):257-265.

[12]He, Y., Ma, W., Li, Y., et al., 2014. Expression of metallothionein of freshwater crab (Sinopotamon henanense) in Escherichia coli enhances tolerance and accumulation of zinc, copper and cadmium. Ecotoxicology, 23(1):56-64.

[13]Jin, R.Y., Gui, W.J., Wu, J.X., et al., 2006. Study on the monoclonal antibody to chlorpyrifos. J. Zhejiang Univ. (Agric. Life Sci.), 32(2):591-597 (in Chinese).

[14]Jin, R.Y., Gui, W.J., Guo, Y.R., et al., 2008. Comparison of monoclonal antibody-based ELISA for triazophos between the indirect and direct formats. Food Agric. Immunol., 19(1):49-60.

[15]Kim, Y.A., Lee, E.H., Kim, K.O., et al., 2011. Competitive immunochromatographic assay for the detection of the organophosphorus pesticide chlorpyrifos. Anal. Chim. Acta, 693(1-2):106-113.

[16]Kim, Y.J., Cho, Y.A., Lee, H., et al., 2003. Investigation of the effect of hapten heterology on immunoassay sensitivity and development of an enzyme-linked immunosorbent assay for the organophosphorus insecticide fenthion. Anal. Chim. Acta, 494(1-2):29-40.

[17]Li, X., Zhang, H., Ji, Y., et al., 2015. Immunochemical and molecular characteristics of monoclonal antibodies against organophosphorus pesticides and effect of hapten structures on immunoassay selectivity. Food Agric. Immunol., 26(1):109-119.

[18]Liang, X., Xie, R., Wang, C., et al., 2013. Development of a broad-selective immunoassay for multi-residue determination of type II pyrethroids in West Lake water. Food Agric. Immunol., 24(1):59-78.

[19]Liu, B.H., Hung, C.T., Lu, C.C., et al., 2014. Production of monoclonal antibody for okadaic acid and its utilization in an ultrasensitive enzyme-linked immunosorbent assay and one-step immunochromatographic strip. J. Agric. Food Chem., 62(6):1254-1260.

[20]Liu, Y.H., Jin, M.J., Gui, W.J., et al., 2007. Hapten design and indirect competitive immunoassay for parathion determination: correlation with molecular modeling and principal component analysis. Anal. Chim. Acta, 591(2):173-182.

[21]Liu, Y.H., Chen, J., Guo, Y.R., et al., 2011. A sensitive monoclonal antibody-based enzyme-linked immunosorbent assay for chlorpyrifos residue determination in Chinese agricultural samples. J. Environ. Sci. Health B, 46(4):313-320.

[22]Lu, S., Zhou, Y., Li, Y., et al., 2012. Production of monoclonal antibody and application in indirect competitive ELISA for detecting okadaic acid and dinophytoxin-1 in seafood. Environ. Sci. Pollut. Res. Int., 19(7):2619-2626.

[23]Manclús, J.J., Primo, J., Montoya, A., 1996. Development of enzyme-linked immunosorbent assays for the insecticide chlorpyrifos. 1. Monoclonal antibody production and immunoassay design. J. Agric. Food Chem., 44(12):4052-4062.

[24]Maquieira, Á., Brun, E.M., Garcés-García, M., et al., 2012. Aluminum oxide nanoparticles as carriers and adjuvants for eliciting antibodies from non-immunogenic haptens. Anal. Chem., 84(21):9340-9348.

[25]Parra, J., Abad-Somovilla, A., Mercader, J.V., et al., 2013. Carbon nanotube-protein carriers enhance size-dependent self-adjuvant antibody response to haptens. J. Control. Release, 170(2):242-251.

[26]Ruan, X., Huang, S., Shao, L., et al., 2013. Monoclonal antibodies against NS4B protein of Japanese encephalitis virus. Monoclon. Antib. Immunodiagn. Immunother., 32(6):382-385.

[27]Stewart, L.D., Elliott, C.T., Walker, A.D., et al., 2009. Development of a monoclonal antibody binding okadaic acid and dinophysistoxins-1, -2 in proportion to their toxicity equivalence factors. Toxicon, 54(4):491-498.

[28]Stils, H.F., 2005. Adjuvants and antibody production: dispelling the myths associated with Freund’s complete and other adjuvants. ILAR J., 46(3):280-293.

[29]Wang, C., Liu, Y., Guo, Y., et al., 2009. Development of a McAb-based immunoassay for parathion and influence of the competitor structure. Food Chem., 115(1):365-370.

[30]Wang, M., Wang, L., Guo, Y., et al., 2015. The broad pattern recognition spectrum of the Toll-like receptor in mollusk Zhikong scallop Chlamys farreri. Dev. Comp. Immunol., 52(2):192-201.

[31]Wang, Z., Zhang, J., Zhang, S., et al., 2011. Heterologous structure of coating antigen on sensitivity of ELISA for sulfamethazine: evidence from molecular similarity analysis. Food Agric. Immunol., 22(2):115-124.

[32]Wongtangprasert, T., Natakuathung, W., Pimpitak, U., et al., 2014. Production of a monoclonal antibody against oxytetracycline and its application for oxytetracycline residue detection in shrimp. J. Zhejiang Univ.-Sci. B (Biomed. & Biotechnol.), 15(2):165-172.

[33]Zhang, D., Li, P., Zhang, Q., et al., 2009. Production of ultrasensitive generic monoclonal antibodies against major aflatoxins using a modified two-step screening procedure. Anal. Chim. Acta, 636(1):63-69.

[34]Zhang, X., Feng, M., Liu, L., et al., 2013a. Detection of aflatoxins in tea samples based on a class-specific monoclonal antibody. Int. J. Food Sci. Technol., 48(6):1269-1274.

[35]Zhang, X., Liu, L., Chen, X., et al., 2013b. Immunochromatographic strip development for ultrasensitive analysis of aflatoxin M1. Anal. Methods, 5(23):6567-6571.

[36]Zhang, X., Zhang, Y., Liu, J., et al., 2013c. PotD protein stimulates biofilm formation by Escherichia coli. Biotechnol. Lett., 35(7):1099-1106.

[37]Zhao, M., Bai, Y., Liu, W., et al., 2013. Immunization of N terminus of enterovirus 71 VP4 elicits cross-protective antibody responses. BMC Microbiol., 13(1):287.

[38]Zhao, Z., Li, B., Sun, H., et al., 2014. Fine-mapping of immunodominant linear B-cell epitopes of the Staphylococcus aureus SEB antigen using short overlapping peptides. PLoS ONE, 9(3):e90445.

Open peer comments: Debate/Discuss/Question/Opinion


Please provide your name, email address and a comment

Journal of Zhejiang University-SCIENCE, 38 Zheda Road, Hangzhou 310027, China
Tel: +86-571-87952783; E-mail: cjzhang@zju.edu.cn
Copyright © 2000 - 2024 Journal of Zhejiang University-SCIENCE