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学者姓名：王荣智
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Abstract ：
Furosine, a hazardous compound mainly produced by the Maillard reaction in food at high temperatures, has been shown to exert cytotoxic effects on cells and cause liver damage after excessive consumption. Hence, it is desirable to develop correlative sensitive and robust immunoassays for furosine detection in real-world samples. In this study, Ovalbumin-furosine (OVA-furo) conjugates were employed as the immunogens for animal immunization, and a hybridoma (1C3) secreting an anti-furosine monoclonal antibody (mAb) was successfully screened. The purified 1C3 mAb exhibited high specificity and affinity (7.4 x 108 L/mol), and an indirect competitive ELISA (ic-ELISA) based on the 1C3 mAb was developed for the detection of furosine. The linear detection range of the ic-ELISA was 6.03-817.85 ng/mL, with a limit of detection (LOD) value of 2 ng/mL. Colloid gold nanospheres(AuNP-) and nanoflower gold-particles(AuNF)-based strips were also established based on this antibody, which had qLOD values and linear ranges of 40 and 20 ng/mL and 40-140 and 20-90 ng/mL, respectively. All the immunoassays showed good detection performance, indicating that these immunoassays have the potential for practical application for the inexpensive detection of furosine in real-world samples.
Keyword ：
ELISA ELISA Furosine Furosine Gold nanoparticles Gold nanoparticles Immunochromatographic strip Immunochromatographic strip Monoclonal antibody Monoclonal antibody
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| GB/T 7714 | Yang, Minyi , Jia, Rongye , Liu, Yuxuan et al. High-sensitivity and rapid immunoassays for furosine detection based on monoclonal antibody 1C3 [J]. | INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES , 2025 , 309 . |
| MLA | Yang, Minyi et al. "High-sensitivity and rapid immunoassays for furosine detection based on monoclonal antibody 1C3" . | INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES 309 (2025) . |
| APA | Yang, Minyi , Jia, Rongye , Liu, Yuxuan , Tang, Hengkun , Wu, Huijuan , Yuan, Jun et al. High-sensitivity and rapid immunoassays for furosine detection based on monoclonal antibody 1C3 . | INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES , 2025 , 309 . |
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mu -KIIIA-Conotoxin (KIIIA) is a short, toxic peptide that selectively targets voltage-gated sodium channels, and has enormous potential in analgesic-drugs development and neuroscience research. However, no correlated immunoassays have been reported for identification and detection of KIIIA. Herein, a hybridoma 3E11 that specifically targets KIIIA was screened using hybridoma technology after animal immunization. The subtype of monoclonal antibody(mAb) 3E11 was IgG1, and it exhibited a high affinity constant (Kaff) of 5.838 x 108 L/mol. Meanwhile, the 3D structure of variable regions of mAb 3E11 was modeled, and the detailed molecular recognition mechanisms of mAb 3E11 to KIIIA were further investigated by molecular docking, alanine scanning and disulfide bond quenching. The sequence "KWCRDH" of KIIIA has been identified as the crucial and structural dependent epitope region recognized by mAb 3E11. The principal forces maintaining the interaction are hydrogen bonding, it-it stacking, nonpolar interactions and salt bridges. Consequently, mAb 3E11 exhibited different binding affinities towards epitope-similar antigens from mu -conotoxin family, including SIIIA, CIIIA, CnIIIA, MIIIA and SmIIIA. Among these mu -conotoxins, the binding affinity of mAb 3E11 to SIIIA is nearly equivalent to that observed with KIIIA. Ultimately, an indirect competitive ELISA(ic-ELISA) was developed based on mAb 3E11, and the linear range of ic-ELISA was 0.72 to 33.02 ng/mL with a lower detection limit (LOD) of 0.28 ng/mL. The recovery rates of intra-assays and inter-assays in spiked samples were 101.32 % and 102.47 %, respectively. The developed ic-ELISA demonstrated high accuracy and repeatability, indicating its potential for detecting the content of KIIIA in real samples.
Keyword ：
Epitope Epitope Immunoassay Immunoassay Molecular recognition Molecular recognition Monoclonal antibody Monoclonal antibody mu-KIIIA-Conotoxin mu-KIIIA-Conotoxin
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| GB/T 7714 | Tang, Hengkun , Wang, Qing , Yang, Minyi et al. Development of sensitive immunoassay for identification and detection of μ-KIIIA-CTX: Insights into antibody discovery, molecular recognition, and immunoassay [J]. | INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES , 2025 , 310 . |
| MLA | Tang, Hengkun et al. "Development of sensitive immunoassay for identification and detection of μ-KIIIA-CTX: Insights into antibody discovery, molecular recognition, and immunoassay" . | INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES 310 (2025) . |
| APA | Tang, Hengkun , Wang, Qing , Yang, Minyi , Jia, Rongye , Yuan, Jun , Wang, Rongzhi . Development of sensitive immunoassay for identification and detection of μ-KIIIA-CTX: Insights into antibody discovery, molecular recognition, and immunoassay . | INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES , 2025 , 310 . |
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Abstract ：
mu -conotoxin KIIIA (KIIIA), as a subtype-specific inhibitor of sodium channel, has demonstrated remarkably potent analgesic properties. Given its significant potential in pharmaceutical development and neuroscience research, it is crucial to establish rapid, effective, and sensitive assay methods for the detection of KIIIA in standardized conotoxin production processes or actual samples. In the present study, colloidal gold nanoparticles (AuNPs), gold nanoflowers (AuNFs) and latex microspheres (LMs) were employed as signal reporters to synthesize immuno-probes through antibody conjugation, facilitating the development of immunochromatographic strips with specificity and accuracy. The sensitivity and specificity of these strips were verified through rigorous experimental procedures, ensuring accurate detection of KIIIA with high feasibility and reliability. The limits of detection (LODs) were determined to be 0.625, 0.078, and 0.156 mu g/mL, with the corresponding visual limits of detection (vLODs) were 10, 5, and 1.25 mu g/mL, respectively. Furthermore, the simplicity and rapidness of the test strips render them a promising tool for on-site monitoring and screening of conotoxin samples, which is crucial for ensuring the safety and efficacy of conotoxin-based biologics. In summary, the three established methods could be applied to detect and analyze KIIIA in conotoxin sample solutions and real samples with excellent sensitivity, portability, and on-site detection. The developed immunoassay method not only fills a gap in the field of conotoxin analysis but also paves the way for future advancements in the quality control and characterization of conotoxin biologics.
Keyword ：
Colloidal gold nanoparticles Colloidal gold nanoparticles Gold nanoflowers Gold nanoflowers Immunochromatographic strip Immunochromatographic strip Latex microspheres Latex microspheres mu-conotoxin KIIIA mu-conotoxin KIIIA
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| GB/T 7714 | Tang, Hengkun , Wang, Qing , Jia, Rongye et al. Development of immunochromatographic strips based on nanoparticle coupled antibody probes for sensitive detection of μ-conotoxin KIIIA [J]. | MICROCHEMICAL JOURNAL , 2025 , 218 . |
| MLA | Tang, Hengkun et al. "Development of immunochromatographic strips based on nanoparticle coupled antibody probes for sensitive detection of μ-conotoxin KIIIA" . | MICROCHEMICAL JOURNAL 218 (2025) . |
| APA | Tang, Hengkun , Wang, Qing , Jia, Rongye , Yuan, Jun , Wang, Rongzhi . Development of immunochromatographic strips based on nanoparticle coupled antibody probes for sensitive detection of μ-conotoxin KIIIA . | MICROCHEMICAL JOURNAL , 2025 , 218 . |
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Abstract ：
采用甲醛法制备了糠氨酸完全抗原，通过交叉验证确保了多克隆抗体的特异性，从而建立了基于该抗体的糠氨酸间接竞争酶联免疫吸附测定（indirect competitive enzyme-linked immunosorbent assay, ic-ELISA）检测方法。结果显示，建立的ic-ELISA检测方法有效检测范围为12.20～2 143.42 ng/mL，最低检测限为2.69 ng/mL。该方法操作简便、迅速，成本低廉，同时具备高灵敏度和优异特异性，为高效检测食品中糠氨酸质量浓度提供了新的选择，具有一定实际应用价值。
Keyword ：
ic-ELISA ic-ELISA 动物免疫 动物免疫 多克隆抗体 多克隆抗体 糠氨酸 糠氨酸
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| GB/T 7714 | 杨敏仪 , 罗雯倩 , 贾荣业 et al. 基于多克隆抗体的糠氨酸ic-ELISA检测方法的建立 [J]. | 食品与生物技术学报 , 2024 , 43 (05) : 138-145 . |
| MLA | 杨敏仪 et al. "基于多克隆抗体的糠氨酸ic-ELISA检测方法的建立" . | 食品与生物技术学报 43 . 05 (2024) : 138-145 . |
| APA | 杨敏仪 , 罗雯倩 , 贾荣业 , 刘宇轩 , 王荣智 . 基于多克隆抗体的糠氨酸ic-ELISA检测方法的建立 . | 食品与生物技术学报 , 2024 , 43 (05) , 138-145 . |
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本发明具体涉及一株能分泌糠氨酸单克隆抗体的杂交瘤细胞株1C3。所述杂交瘤细胞株1C3的分类命名为抗糠氨酸单克隆抗体杂交瘤细胞株，其已于2023年10月10日在中国普通微生物菌种保藏管理中心登记保藏，保藏编号为CGMCC NO.45728。杂交瘤细胞株1C3分泌的抗糠氨酸单克隆抗体特异性强、亲和力大、效价高。基于杂交瘤细胞株1C3及其所分泌的单克隆抗体，本发明建立了一种ELISA免疫检测方法，该检测方法可应用于牛奶中糠氨酸的检测，且具有准确性强、灵敏度高、操作简便的优势。
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| GB/T 7714 | 王荣智 , 杨敏仪 , 贾荣业 et al. 一株能稳定分泌抗糠氨酸单克隆抗体的杂交瘤细胞株 : CN202311833979.6[P]. | 2023-12-28 . |
| MLA | 王荣智 et al. "一株能稳定分泌抗糠氨酸单克隆抗体的杂交瘤细胞株" : CN202311833979.6. | 2023-12-28 . |
| APA | 王荣智 , 杨敏仪 , 贾荣业 , 刘宇轩 , 吴惠娟 , 罗雯倩 et al. 一株能稳定分泌抗糠氨酸单克隆抗体的杂交瘤细胞株 : CN202311833979.6. | 2023-12-28 . |
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Abstract ：
本发明提供一种检测河豚毒素的乳胶微球免疫层析试纸条及其应用，属于海洋毒素快速检测领域。所述乳胶微球免疫层析试纸条包括以下组分：塑料外壳、样品垫、免疫探针结合垫、硝酸纤维膜和吸水垫；所述免疫探针结合垫滴加有抗河豚毒素单克隆抗体标记的乳胶微球免疫探针；所述抗河豚毒素单克隆抗体由抗河豚毒素单克隆抗体杂交瘤细胞株5B9分泌，杂交瘤细胞株5B9保藏号为CGMCC No.45320。该乳胶微球免疫层析试纸条，检测阈值为1000ng/mL，检测限为7.8125ng/mL，特异性强，灵敏度高，重复性良好，性能稳定，操作要求低，非常适用于食品中快速检测，对监测河豚毒素具有重要意义。
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| GB/T 7714 | 凌素美 , 黄永铭 , 汪世华 et al. 一种检测河豚毒素的乳胶微球免疫层析试纸条及其应用 : CN202310016526.8[P]. | 2023-01-06 . |
| MLA | 凌素美 et al. "一种检测河豚毒素的乳胶微球免疫层析试纸条及其应用" : CN202310016526.8. | 2023-01-06 . |
| APA | 凌素美 , 黄永铭 , 汪世华 , 徐杨 , 徐爱迪 , 孙梦晗 et al. 一种检测河豚毒素的乳胶微球免疫层析试纸条及其应用 : CN202310016526.8. | 2023-01-06 . |
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Tetrodotoxin (TTX) could result in serious diseases due to its extremely high neurotoxicity. Thus, it is of great importance to measure TTX for food safety. In this study, an anti-TTX monoclonal antibody with good specificity and high affinity was used to develop the immunochromatographic test strips (ICTS). Gold nanoflower (AuNF) with multiple branches and latex microsphere (LM) with large particle size as signal reporters were employed for improving the sensitivity of test strips. Both AuNF and LM probes are stable, and the developed ICTS were specific to TTX, demonstrating no cross-reactivity with other marine toxins. The linear range of AuNF- and LM-based strips for TTX was 9.49-330.98 ng/mL and 5.40-443.19 ng/mL, respectively. The limit of detection (LOD) of AuNF- and LM-based strips was determined to be 9.49 ng/mL and 5.40 ng/mL, respectively. In summary, the developed ICTS based on AuNF and LM signal probes displayed enhancement of sensitivity and provided rapid and specific detection of TTX.
Keyword ：
detection detection gold nanoflower gold nanoflower immunochromatography strip immunochromatography strip latex microsphere latex microsphere monoclonal antibody monoclonal antibody tetrodotoxin tetrodotoxin
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| GB/T 7714 | Huang, Yongming , Xu, Aidi , Xu, Yang et al. Sensitive and rapid detection of tetrodotoxin based on gold nanoflower-and latex microsphere-labeled monoclonal antibodies [J]. | FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY , 2023 , 11 . |
| MLA | Huang, Yongming et al. "Sensitive and rapid detection of tetrodotoxin based on gold nanoflower-and latex microsphere-labeled monoclonal antibodies" . | FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY 11 (2023) . |
| APA | Huang, Yongming , Xu, Aidi , Xu, Yang , Wu, Huijuan , Sun, Menghan , Madushika, Lakshani et al. Sensitive and rapid detection of tetrodotoxin based on gold nanoflower-and latex microsphere-labeled monoclonal antibodies . | FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY , 2023 , 11 . |
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In view of the toxicological hazard and important applications in analgesics and cancer chemotherapeutics of alpha B-CTX, it is urgent to develop an accurate, effective and feasible immunoassay for the determination and analysis of alpha B-CTX in real samples. In this study, MBP-alpha B-CTX4 tandem fusion protein was used as an immunogen to elicit a strong immune response, and a hybridoma cell 5E4 secreting IgG2b against alpha B-CTX was successfully screened by hybridoma technology. The affinity of the purified 5E4 monoclonal antibody (mAb) was 1.02 x 10(8) L/mol, which showed high affinity and specificity to alpha B-CTX. Epitope 1 of alpha B-CTX is the major binding region for 5E4 mAb recongnization, and two amino acid residues (14L and 15F) in alpha B-CTX were critical sites for the interaction between alpha B-CTX and 5E4 mAb. Indirect competitive ELISA (ic-ELISA) based on 5E4 mAb was developed to detect and analyze alpha B-CTX in real samples, and the linear range of ic-ELISA to alpha B-CTX was 117-3798 ng/mL, with a limit of detection (LOD) of 81 ng/mL. All the above results indicated that the developed ic-ELISA had high accuracy and repeatability, and it could be applied for alpha B-CTX detection and drug analysis in real samples.
Keyword ：
alpha B-conotoxin VxXXIVA alpha B-conotoxin VxXXIVA ELISA ELISA epitope epitope hybridoma hybridoma monoclonal antibody monoclonal antibody
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| GB/T 7714 | Tang, Hengkun , Liu, Haimei , Gao, Yehong et al. Detection of αB-Conotoxin VxXXIVA (αB-CTX) by ic-ELISA Based on an Epitope-Specific Monoclonal Antibody [J]. | TOXINS , 2022 , 14 (3) . |
| MLA | Tang, Hengkun et al. "Detection of αB-Conotoxin VxXXIVA (αB-CTX) by ic-ELISA Based on an Epitope-Specific Monoclonal Antibody" . | TOXINS 14 . 3 (2022) . |
| APA | Tang, Hengkun , Liu, Haimei , Gao, Yehong , Chen, Rui , Dong, Mingke , Ling, Sumei et al. Detection of αB-Conotoxin VxXXIVA (αB-CTX) by ic-ELISA Based on an Epitope-Specific Monoclonal Antibody . | TOXINS , 2022 , 14 (3) . |
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Abstract ：
Given the application of alpha B-VxXXIVA-conotoxin (alpha B-CTX) in analgesics and cancer chemotherapeutics, and its threat to humans, it is urgent to develop a rapid, effective and accurate method for the analysis and detection of alpha B-CTX in real shellfish and medicine drug samples. In the present study, two different immunochromatographic strips were established for alpha B-CTX detection, based on the monoclonal antibody 5E4 against alpha B-CTX, and the visual limits of detection (vLOD) for the colloidal gold nanoparticles-based strip (AuNPs-based strip) and nanoflowers-based strip (AuNFs-based strip) were 4 mu g/mL and 1.5 mu g/mL, respectively. The developed AuNPs/AuNFs-based strips have good specificity and accuracy, and the detection results were analyzed in less than 10 min, without using an instrument. In view of the excellent repeatability and usability, the established methods could be applied to detect and analyze the content of alpha B-CTX in real samples.
Keyword ：
alpha B-VxXXIVA-conotoxin alpha B-VxXXIVA-conotoxin AuNFs-based strip AuNFs-based strip AuNPs-based strip AuNPs-based strip detection detection monoclonal antibody monoclonal antibody
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| GB/T 7714 | Tang, Hengkun , Liu, Haimei , Chen, Rui et al. Development of Immunochromatographic Strip for Detection of αB-VxXXIVA-Conotoxin Based on 5E4 Monoclonal Antibody [J]. | TOXINS , 2022 , 14 (3) . |
| MLA | Tang, Hengkun et al. "Development of Immunochromatographic Strip for Detection of αB-VxXXIVA-Conotoxin Based on 5E4 Monoclonal Antibody" . | TOXINS 14 . 3 (2022) . |
| APA | Tang, Hengkun , Liu, Haimei , Chen, Rui , Gao, Yehong , Dong, Mingke , Ling, Sumei et al. Development of Immunochromatographic Strip for Detection of αB-VxXXIVA-Conotoxin Based on 5E4 Monoclonal Antibody . | TOXINS , 2022 , 14 (3) . |
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Tenuazonic acid (TA) is a highly toxic mycotoxin mainly generated by the fungi of Alternaria genus and widely contaminates agricultural by-products. Given the threat of TA to food-security, it is very important to develop rapid and sensitive detection methods for TA monitoring. In this study, gold nano-particles (AuNP) with average diameter near 17.25 nm were prepared, and the developed AuNP-based strip has an assay time of 15 min with visual limit of detection (LOD) of 12.5 ng/ml and threshold of 100 ng/ml. To further improve sensitivity, multi-branched gold nano-flowers (AuNF) with average diameter near 50 nm were prepared and characterized by UV-VIS and TEM, and the established AuNF-based strip has visual LOD of 0.78 ng/ml and threshold of 50 ng/ml within 15 min. Both assays were applied to determine TA in apple juice and tomato ketchup, and the results were consistent with that of UHPLC-MS/MS. Thus, these assays could be applied for rapid determination of trace TA in real samples.
Keyword ：
colloidal gold colloidal gold lateral flow immunochromatographic assay lateral flow immunochromatographic assay multi-branched gold nano-flower particle multi-branched gold nano-flower particle mycotoxin mycotoxin TA TA test strip test strip
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| GB/T 7714 | Cai, Peiyuan , Wang, Rongzhi , Ling, Sumei et al. Rapid and sensitive detection of tenuazonic acid in agricultural by-products based on gold nano-flower lateral flow [J]. | FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY , 2022 , 10 . |
| MLA | Cai, Peiyuan et al. "Rapid and sensitive detection of tenuazonic acid in agricultural by-products based on gold nano-flower lateral flow" . | FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY 10 (2022) . |
| APA | Cai, Peiyuan , Wang, Rongzhi , Ling, Sumei , Wang, Shihua . Rapid and sensitive detection of tenuazonic acid in agricultural by-products based on gold nano-flower lateral flow . | FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY , 2022 , 10 . |
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