抗独特型抗体在半抗原免疫检测中的应用

doi:10.19586/j.2095-2341.2023.0066

生物技术进展 ›› 2023, Vol. 13 ›› Issue (5): 690-697.DOI: 10.19586/j.2095-2341.2023.0066

抗独特型抗体在半抗原免疫检测中的应用

杜凯(), 张卓玲, 高莉, 何海华, 李婷华, 饶微()

深圳市新产业生物医学工程股份有限公司，广东深圳 518118

收稿日期:2023-05-12 接受日期:2023-06-09 出版日期:2023-09-25 发布日期:2023-10-10
通讯作者: 饶微
作者简介:杜凯 E-mail: kai.du@snibe.cn；
基金资助:
深圳市科技创新委员会科技抗疫专项(JSGG20220919091201003)

Application of Anti-idiotype Antibodies in Haptens Immunoassay

Kai DU(), Zhuoling ZHANG, Li GAO, Haihua HE, Tinghua LI, Wei RAO()

Shenzhen New Industries Biomedical Engineering Co. ，Ltd，Guangdong Shenzhen 518118，China

Received:2023-05-12 Accepted:2023-06-09 Online:2023-09-25 Published:2023-10-10
Contact: Wei RAO

摘要/Abstract

摘要：

小分子半抗原的检测在食品药品检验、环境检测和疾病诊断等领域具有重要意义，但目前针对半抗原的免疫检测在灵敏度方面表现欠佳。抗独特型抗体（anti-idiotypes，Ab2）发现于20世纪初，主要分为3种亚型：Ab2α、Ab2β和Ab2γ。Ab2除了可以用于预防和治疗疾病外，在免疫检测中也有潜在的应用价值。概述了Ab2在竞争性、非竞争性及基于噬菌体免疫分析中的应用，包括检测模式及效果。无论是在竞争法中采用合适的Ab2替代半抗原衍生物，还是在非竞争法中采用针对一抗和半抗原复合物的抗伴型抗体，都能提高检测灵敏度，改善交叉率。Ab2的应用是未来体外诊断试剂盒开发的一个重要思路。此外，还对目前Ab2的几种开发方法进行了分析，较之传统抗体开发技术，天然纳米抗体库可能是获取Ab2更为实用的一种方式。

关键词: 抗独特型抗体, 半抗原, 免疫分析, 纳米抗体库

Abstract:

The detection of small molecule haptens is of great significance for food and drug testing， environmental monitoring and disease diagnosis. However， commercially available immunoassays for haptens are limited， partially due to poor performance， such as low sensitivity. In the early 20th century， anti-idiotypes （Ab2） were discovered， and Ab2 can be divided into three different isoforms： Ab2α， Ab2β and Ab2γ. In addition to be used for the treatment and prevention of disease， Ab2 has potential applications in immunoassays. This review provided an overview of the use of Ab2 in competitive， non-competitive and phage-based immunoassays， including the different detection formats and effects on the performance. Both the use of appropriate Ab2 instead of hapten derivatives in competitive assays and the use of anti-metatype antibodies against primary antibody and hapten complexes in non-competitive assays could increase detection sensitivity and improve cross-reactivity. The use of Ab2 could be a potentially important choice for the development of in vitro diagnostic assays in the future. In addition， different methods for the generation of Ab2 were summarized. The selection of natural nanobody libraries might be a more practical way to obtain Ab2 than traditional monoclonal antibody production.

Key words: anti-idiotypes, hapten, immunoassay, nanobody library

中图分类号:

Q503

杜凯, 张卓玲, 高莉, 何海华, 李婷华, 饶微. 抗独特型抗体在半抗原免疫检测中的应用[J]. 生物技术进展, 2023, 13(5): 690-697.

Kai DU, Zhuoling ZHANG, Li GAO, Haihua HE, Tinghua LI, Wei RAO. Application of Anti-idiotype Antibodies in Haptens Immunoassay[J]. Current Biotechnology, 2023, 13(5): 690-697.

图/表 7

图1 Ab2的分类[8]

Fig. 1 The classification of Ab2[8]

图2 免疫层析竞争法检测真菌毒素的2种模式A：单克隆抗体包被纳米球；B：单域重链抗体包被纳米球[22]。C—质控线，T—检测线。

Fig. 2 Two patterns of the competitive immunochromatographic assays for mycotoxin

图3 基于Ab2的非竞争检测模式[31]

Fig. 3 Schematic illustration of Ab2-based noncompetitive immunoassay format[31]

图4 免疫层析系统检测[34]A：免疫层析系统的组成；B：无分析物时不显色；C：有分析物时显色

Fig. 4 Detection of the immunochromatography system[34]

表1 非竞争法与竞争法分析性能指标对比数据

Table 1 Comparison of performance indicators between non-competitive and competitive assays

指标	E2/(pg·mL^-1)		25-OH VD/(ng·mL^-1)		ALD/(pg·mL^-1)
指标	竞争法	非竞争法	竞争法	非竞争法	竞争法	非竞争法
空白限	5.0	1.0	3.0	0.05	5.0	5.0
检测限	12.0	5.0	4.5	0.50	10.0	8.0
测量范围	5~6 000	1~48 000	3.0~150	0.05~300	5~1 000	5~2 000
线性范围	100~3 000	10~4 800	10~150	1.5~150	20~1 000	30~2 000

图5 抗伴型抗体的结合位点

Fig. 5 The binding site of anti-metatype antibody

图6 免疫环介导等温扩增的原理[44]

Fig. 6 Schematic diagram of the immuno-loop-mediated isothermal amplification (iLAMP)[44].

参考文献 44

1	KARASHIMA S, OSAKA I. Rapidity and precision of steroid hormone measurement[J/OL]. J. Clin. Med., 2022, 11(4): 956[2023-07-22]. .
2	XIONG Y, LENG Y, LI X, et al.. Emerging strategies to enhance the sensitivity of competitive ELISA for detection of chemical contaminants in food samples[J/OL]. Trac Trends Anal. Chem., 2020, 126: 115861[2023-07-22]. .
3	雷豆豆, 马润然, 王伽伯, 等. 食品中农药残留的新型生物检测技术研究进展[J]. 生物技术进展, 2023, 13(1): 1-10.
4	TSAGKARIS A S, NELIS J L D, ROSS G M S, et al.. Critical assessment of recent trends related to screening and confirmatory analytical methods for selected food contaminants and allergens[J/OL]. Trac Trends Anal. Chem., 2019, 121: 115688[2023-07-22]. .
5	LI Y, ZHANG G, MAO X, et al.. High sensitivity immunoassays for small molecule compounds detection-novel noncompetitive immunoassay designs[J]. Trac Trends Anal. Chem., 2018, 103: 198-208.
6	ZHAO F, SHI R, LIU R, et al.. Application of phage-display developed antibody and antigen substitutes in immunoassays for small molecule contaminants analysis: a mini-review[J/OL]. Food Chem., 2021, 339: 128084[2023-07-22]. .
7	LI P, DENG S, ZECH XU Z. Toxicant substitutes in immunological assays for mycotoxins detection: a mini review[J/OL]. Food Chem., 2021, 344: 128589[2023-07-22]. .
8	PAN S Y, CHIA Y C, YE E, et al.. Immunomodulatory potential of anti-idiotypic antibodies for the treatment of autoimmune diseases[J/OL]. Future Sci., 2020, 7(2): FSO648[2023-07-22]. .
9	STANOVA A K, RYABKOVA V A, UTEKHIN S V, et al.. Anti-idiotypic agonistic antibodies: candidates for the role of universal remedy[J/OL]. Antibodies, 2020, 9(2): 19[2023-07-22]. .
10	MURPHY W J, LONGO D L. A possible role for anti-idiotype antibodies in SARS-CoV-2 infection and vaccination[J]. N. Engl. J. Med., 2022, 386(4): 394-396.
11	SPINKS C A, WANG B, MILLS E N C, et al.. Production and characterization of monoclonal anti‐idiotype antibody mimics for the pyrethroid insecticides and the herbicide paraquat[J]. Food Agric. Immunol., 1993, 5(1): 13-25.
12	CHEN L, HU X, XING Y, et al.. Highly sensitive immunochromatographic assay for simultaneous determination of azaperone and azaperol in pork[J/OL]. Food Chem. X, 2023, 17: 100525[2023-07-22]. .
13	LIAO J, ZHUO X, PAN B, et al.. Synthesis and preliminary immunologic properties of di-/ trisaccharide-conjugates related to Bacillus anthracis [J/OL]. Bioorg. Med. Chem. Lett., 2022, 76: 128986[2023-07-22]. .
14	JONES W, HARVEY D, MITCHELL R. Production of anti-idiotypic monoclonal antibody mimics for coronatine[J]. Food Agric. Immunol., 2002, 14(4): 301-311.
15	JONES W T, HARVEY D, SUTHERLAND P W, et al.. Production of anti-idiotypic monoclonal antibodies that mimic the phytotoxin dothistromin[J]. Food Agric. Immunol., 1998, 10(1): 67-78.
16	高莉, 杨丽芳, 邱仁东, 等. 一种特异性结合 E2-E2抗体复合物的抗体及其应用: CN116041527A[P]. 2023-05-02.
17	KIRSCH R D, BEALE D, HE M, et al.. Anti-anti-idiotypic (Ab3) antibodies that bind progesterone-11alpha-bovine serum albumin differ in their combining sites from antibodies raised directly against the antigen[J]. Immunology, 2000, 100(2): 152-164.
18	SHI L, YU T, LUO M, et al.. Preparation monoclonal β-type anti-idiotype antibody of zearalenone and development of green ELISA quantitative detecting technique[J]. Prep. Biochem. Biotechnol., 2020, 50(4): 419-424.
19	ZHANG C, ZHANG Q, TANG X, et al.. Development of an anti-idiotypic VHH antibody and toxin-free enzyme immunoassay for ochratoxin A in cereals[J/OL]. Toxins, 2019, 11(5): 280[2023-07-22]. .
20	HU L, LIU A, CHEN W, et al.. A non-toxic enzyme-linked immunosorbent assay for aflatoxin B₁ using anti-idiotypic antibodies as substitutes[J]. J. Sci. Food Agric., 2017, 97(5): 1640-1645.
21	SCHULZ K, PÖHLMANN C, DIETRICH R, et al.. Electrochemical biochip assays based on anti-idiotypic antibodies for rapid and automated on-site detection of low molecular weight toxins[J/OL]. Front. Chem., 2019, 7: 31[2023-07-22]. .
22	TANG X, LI P, ZHANG Q, et al.. Time-resolved fluorescence immunochromatographic assay developed using two idiotypic nanobodies for rapid, quantitative, and simultaneous detection of aflatoxin and zearalenone in maize and its products[J]. Anal. Chem., 2017, 89(21): 11520-11528.
23	AKKOYUN A, KOHEN V F, BILITEWSKI U. Detection of sulphamethazine with an optical biosensor and anti-idiotypic antibodies[J]. Sens. Actuat. B Chem., 2000, 70(1-3): 12-18.
24	SCHULZ K, PÖHLMANN C, DIETRICH R, et al.. An electrochemical fiveplex biochip assay based on anti-idiotypic antibodies for fast on-site detection of bioterrorism relevant low molecular weight toxins[J/OL]. Toxins, 2019, 11(12): 696[2023-07-22]. .
25	XU Y, XIONG L, LI Y, et al.. Anti-idiotypic nanobody as citrinin mimotope from a naive alpaca heavy chain single domain antibody library[J]. Anal. Bioanal. Chem., 2015, 407(18): 5333-5341.
26	SHU M, XU Y, WANG D, et al.. Anti-idiotypic nanobody: a strategy for development of sensitive and green immunoassay for fumonisin B₁[J]. Talanta, 2015, 143: 388-393.
27	ZHANG C, ZHANG W, TANG X, et al.. Change of amino acid residues in idiotypic nanobodies enhanced the sensitivity of competitive enzyme immunoassay for mycotoxin ochratoxin A in cereals[J/OL]. Toxins, 2020, 12(4): 273[2023-07-22]. .
28	QIU Y L, HE Q H, XU Y, et al.. Deoxynivalenol-mimic nanobody isolated from a naïve phage display nanobody library and its application in immunoassay[J]. Anal. Chim. Acta, 2015, 887: 201-208.
29	QIU Y L, HE Q H, XU Y, et al.. Modification of a deoxynivalenol-antigen-mimicking nanobody to improve immunoassay sensitivity by site-saturation mutagenesis[J]. Anal. Bioanal. Chem., 2016, 408(3): 895-903.
30	XIONG L, ZHANG X, XU Y, et al.. Anti-idiotypic VHH mediated environmentally friendly immunoassay for citrinin without mycotoxin[J]. Food Agric. Immunol., 2020, 31(1): 968-984.
31	MILLS C, CAMPBELL K. A new chapter for anti-idiotypes in low molecular weight compound immunoassays[J]. Trends Biotechnol., 2022, 40(9): 1102-1120.
32	KOBAYASHI N, IWAKAMI K, KOTOSHIBA S, et al.. Immunoenzymometric assay for a small molecule, 11-deoxycortisol, with attomole-range sensitivity employing an scFv-enzyme fusion protein and anti-idiotype antibodies[J]. Anal. Chem., 2006, 78(7): 2244-2253.
33	KOBAYASHI N, OIWA H, KUBOTA K, et al.. Monoclonal antibodies generated against an affinity-labeled immune complex of an anti-bile acid metabolite antibody: an approach to noncompetitive hapten immunoassays based on anti-idiotype or anti-metatype antibodies[J]. J. Immunol. Meth., 2000, 245(1-2): 95-108.
34	WILD D. The immunoassay handbook: theory and applications of ligand binding, ELISA and related techniques[M]. USA, The Netherlands: Elsevier Science, 2013.
35	OMI K, ANDO T, SAKYU T, et al.. Noncompetitive immunoassay detection system for haptens on the basis of antimetatype antibodies[J]. Clin. Chem., 2015, 61(4): 627-635.
36	SHU M, XU Y, DONG J X, et al.. Development of a noncompetitive idiometric nanobodies phage immumoassay for the determination of fumonisin B₁ [J]. Food Agric. Immunol., 2019, 30(1): 510-521.
37	LI X, LI P, ZHANG Q, et al.. Molecular characterization of monoclonal antibodies against aflatoxins: a possible explanation for the highest sensitivity[J]. Anal. Chem., 2012, 84(12): 5229-5235.
38	WANG Y, LI P, MAJKOVA Z, et al.. Isolation of alpaca anti-idiotypic heavy-chain single-domain antibody for the aflatoxin immunoassay[J]. Anal. Chem., 2013, 85(17): 8298-8303.
39	ZHAO Y, LIANG Y, LIU Y, et al.. Phage displayed domain antibody mimics for pyrethroid and its application in immunoassay[J]. Int. J. Agric. Biol. Eng., 2020, 13(2): 235-240.
40	JI Y, HE Q, XU Y, et al.. Phage displayed anti-idiotypic nanobody mediated immuno-PCR for sensitive and environmentally friendly detection of mycotoxin ochratoxin A[J]. Anal. Methods, 2016, 8(43): 7824-7831.
41	HUANG W, TU Z, NING Z, et al.. Development of real-time immuno-PCR based on phage displayed an anti-idiotypic nanobody for quantitative determination of citrinin in Monascus [J/OL]. Toxins, 2019, 11(10): 572[2023-07-23]. .
42	WANG X, HE Q, XU Y, et al.. Anti-idiotypic VHH phage display-mediated immuno-PCR for ultrasensitive determination of mycotoxin zearalenone in cereals[J]. Talanta, 2016, 147: 410-415.
43	LEI J, LI P, ZHANG Q, et al.. Anti-idiotypic nanobody-phage based real-time immuno-PCR for detection of hepatocarcinogen aflatoxin in grains and feedstuffs[J]. Anal. Chem., 2014, 86(21): 10841-10846.
44	LEI J, HAN X, TANG X, et al.. Development of anti-idiotypic nanobody-phage based immuno-loop-mediated isothermal amplification assay for aflatoxins in peanuts[J/OL]. Toxins, 2020, 12(9): 565[2023-07-23]. .

[1]	雷豆豆, 马润然, 王伽伯, 孔维军. 食品中农药残留的新型生物检测技术研究进展[J]. 生物技术进展, 2023, 13(1): 1-10.
[2]	薛虎寅,尹永梅,张太昌,宋佩,田溪,张昱,孟萌,郗日沫. 偶氮类合成色素检测技术的研究进展[J]. 生物技术进展, 2012, 2(3): 171-176.

抗独特型抗体在半抗原免疫检测中的应用

Application of Anti-idiotype Antibodies in Haptens Immunoassay

RichHTML

PDF

可视化

摘要/Abstract

引用本文

使用本文

图/表 7

参考文献 44

相关文章 2

编辑推荐

Metrics

本文评价