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"intrinsically unstructured domain"

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"intrinsically unstructured domain"

Brief Communication

Western Blot Detection of Human Anti-Chikungunya Virus Antibody with Recombinant Envelope 2 Protein
Zhaoshou Yang, Jihoo Lee, Hye-Jin Ahn, Chom-Kyu Chong, Ronaldo F. Dias, Ho-Woo Nam
Korean J Parasitol 2016;54(2):239-241.
Published online April 30, 2016
DOI: https://doi.org/10.3347/kjp.2016.54.2.239
Chikungunya virus (CHIKV), a tropical pathogen, has re-emerged and has massive outbreaks abruptly all over the world. Containing many dominant epitopes, the envelope E2 protein of CHIKV has been explored for the vaccination or diagnosis. In the present study, the antigenicity of a recombinant expressed intrinsically disorder domain (IUD) of E2 was tested for the detection of the antibody against CHIKV through western blot method. The gene of the IUD of E2 was inserted into 2 different vectors and expressed as recombinant GST-E2 and recombinant MBP-E2 fusion protein, respectively. Two kinds of fusion proteins were tested with 30 CHIKV patient sera and 30 normal sera, respectively. Both proteins were detected by 25 patients sera (83.3%) and 1 normal serum (3.3%). This test showed a relatively high sensitivity and very high specificity of the recombinant E2 proteins to be used as diagnostic antigens against CHIKV infection.

Citations

Citations to this article as recorded by  Crossref logo
  • Medical viruses: diagnostic techniques
    Harsh, Pratima Tripathi
    Virology Journal.2023;[Epub]     CrossRef
  • Diagnostic accuracy of serological tests for the diagnosis of Chikungunya virus infection: A systematic review and meta-analysis
    Anna Andrew, Tholasi Nadhan Navien, Tzi Shien Yeoh, Marimuthu Citartan, Ernest Mangantig, Magdline S. H. Sum, Ewe Seng Ch’ng, Thean-Hock Tang, Guilherme S. Ribeiro
    PLOS Neglected Tropical Diseases.2022; 16(2): e0010152.     CrossRef
  • An Overview of Laboratory Diagnosis of Central Nervous System Viral Infections
    Sharifah Aliah Diyanah Syed Hussin, Ang-Lim Chua, Hassanain Al-Talib, Shamala Devi Sekaran, Seok Mui Wang
    Journal of Pure and Applied Microbiology.2022; 16(4): 2225.     CrossRef
  • Molecular and Immunological Diagnostic Techniques of Medical Viruses
    Daniel Hussien Reta, Tesfaye Sisay Tessema, Addis Simachew Ashenef, Adey Feleke Desta, Wajana Lako Labisso, Solomon Tebeje Gizaw, Solomon Mequanente Abay, Daniel Seifu Melka, Fisseha Alemu Reta
    International Journal of Microbiology.2020; 2020: 1.     CrossRef
  • Apigenin enhances the cisplatin cytotoxic effect through p53-modulated apoptosis
    Rui Liu, Ping Ji, Bin Liu, Haishi Qiao, Xia Wang, Likun Zhou, Ting Deng, Yi Ba
    Oncology Letters.2017; 13(2): 1024.     CrossRef
  • 9,202 View
  • 164 Download
  • 6 Web of Science
  • Crossref
Original Articles
High Expression of Water-Soluble Recombinant Antigenic Domains of Toxoplasma gondii Secretory Organelles
Zhaoshou Yang, Hye-Jin Ahn, Ho-Woo Nam
Korean J Parasitol 2014;52(4):367-376.
Published online August 29, 2014
DOI: https://doi.org/10.3347/kjp.2014.52.4.367

Recombinant antigenic proteins of Toxoplasma gondii are alternative source of antigens which are easily obtainable for serodiagnosis of toxoplasmosis. In this study, highly antigenic secretory organellar proteins, dense granular GRA2 and GRA3, rhoptrial ROP2, and micronemal MIC2, were analyzed by bioinformatics approach to express as water-soluble forms of antigenic domains. The transmembrane region and disorder tendency of 4 secretory proteins were predicted to clone the genes into pGEX-4T-1 vector. Recombinant plasmids were transformed into BL21 (DE3) pLysS E. coli, and GST fusion proteins were expressed with IPTG. As a result, GST fusion proteins with GRA225-105, GRA339-138, ROP2324-561, and MIC21-284 domains had respectively higher value of IgG avidity. The rGST-GRA225-105 and rGST-GRA339-138 were soluble, while rGST-ROP2324-561 and rGST-MIC21-284 were not. GRA231-71, intrinsically unstructured domain (IUD) of GRA2, was used as a linker to enhance the solubility. The rGST-GRA231-71-ROP2324-561, a chimeric protein, appeared to be soluble. Moreover, rGST-GRA231-71-MIC21-284 was also soluble and had higher IgG avidity comparing to rGST-MIC21-284. These 4 highly expressed and water-soluble recombinant antigenic proteins may be promising candidates to improve the serodiagnosis of toxoplasmosis in addition to the major surface antigen of SAG1.

Citations

Citations to this article as recorded by  Crossref logo
  • Single Cell Expression Systems for the Production of Recombinant Proteins for Immunodiagnosis and Immunoprophylaxis of Toxoplasmosis
    Karolina Sołowińska, Lucyna Holec-Gąsior
    Microorganisms.2024; 12(8): 1731.     CrossRef
  • A vaccine using Anaplasma marginale subdominant type IV secretion system recombinant proteins was not protective against a virulent challenge
    Macarena Sarli, María B. Novoa, Matilde N. Mazzucco, Marcelo L. Signorini, Ignacio E. Echaide, Susana T. de Echaide, María E. Primo, Paulo Lee Ho
    PLOS ONE.2020; 15(2): e0229301.     CrossRef
  • Development of a novel fusion protein with Anaplasma marginale and A. centrale MSP5 improved performance of Anaplasma antibody detection by cELISA in infected and vaccinated cattle
    María E. Primo, Carolina S. Thompson, Beatriz S. Valentini, Macarena Sarli, María B. Novoa, Atilio J. Mangold, Susana T. de Echaide, Ulrike Gertrud Munderloh
    PLOS ONE.2019; 14(1): e0211149.     CrossRef
  • TheToxoplasma gondiidense granule protein TgGRA3 interacts with host Golgi and dysregulates anterograde transport
    Maika S. Deffieu, Tchilabalo Dilezitoko Alayi, Christian Slomianny, Stanislas Tomavo
    Biology Open.2019;[Epub]     CrossRef
  • 9,699 View
  • 77 Download
  • 4 Web of Science
  • Crossref
A Rapid Diagnostic Test for Toxoplasmosis using Recombinant Antigenic N-terminal Half of SAG1 Linked with Intrinsically Unstructured Domain of GRA2 Protein
Kyoung Ju Song, Zhaoshou Yang, Chom-Kyu Chong, Jin-Soo Kim, Kyung Chan Lee, Tong-Soo Kim, Ho-Woo Nam
Korean J Parasitol 2013;51(5):503-510.
Published online October 31, 2013
DOI: https://doi.org/10.3347/kjp.2013.51.5.503

Toxoplasma gondii is an apicomplexan parasite with a broad host range of most warm-blooded mammals including humans, of which one-thirds of the human population has been infected worldwide which can cause congenital defects, abortion, and neonatal complications. Here, we developed a rapid diagnostic test (RDT) for T. gondii infection. Antigenic N-terminal half of the major surface antigen (SAG1) was linked with intrinsically unstructured domain (IUD) of dense granule protein 2 (GRA2). The recombinant GST-GRA2-SAG1A protein was successfully expressed and purified as 51 kDa of molecular weight. Furthermore, antigenicity and solubility of the rGST-GRA2-SAG1A protein were significantly increased. The overall specificity and sensitivity of GST-GRA2-SAG1A loaded RDT (TgRDT) were estimated as 100% and 97.1% by comparing with ELISA result which uses T. gondii whole cell lysates as the antigen. The TgRDT tested with Uganda people sera for field trial and showed 31.9% of seroprevalence against T. gondii antibody. The TgRDT is proved to be a kit for rapid and easy to use with high accuracy, which would be a suitable serodiagnostic tool for toxoplasmosis.

Citations

Citations to this article as recorded by  Crossref logo
  • Development of an Immunochromatographic Test with Recombinant MIC2-MIC3 Fusion Protein for Serological Detection of Toxoplasma gondii
    Jianzhong Wang, Yi Zhao, Jicheng Qiu, Jing Liu, Rui Zhou, Xialin Ma, Xiaojie Wu, Xiaoguang Li, Wei Mao, Yiduo Liu, Heng Zhang
    Veterinary Sciences.2025; 12(6): 509.     CrossRef
  • Application of gold immunochromatographic assay strip combined with digital evaluation for early detection of Toxoplasma gondii infection in multiple species
    Jiyuan Fan, Hao Sun, Jiawen Fang, Yafan Gao, Haojie Ding, Bin Zheng, Qingming Kong, Xunhui Zhuo, Shaohong Lu
    Parasites & Vectors.2024;[Epub]     CrossRef
  • Single Cell Expression Systems for the Production of Recombinant Proteins for Immunodiagnosis and Immunoprophylaxis of Toxoplasmosis
    Karolina Sołowińska, Lucyna Holec-Gąsior
    Microorganisms.2024; 12(8): 1731.     CrossRef
  • Seroprevalence of Toxoplasma gondii infection among patients of a tertiary hospital in Guangzhou, Guangdong province, PR China
    Yu-bin Guan, Xiao-xiao Sun, Shao-lian Chen, Xiao-ting Zhu, Zhi-hua Zeng, Han-wei Lu, Hong-mei Feng, Yu Guo, Wen-gong Jiang, Kui Xiong, Xiao-rong Yang, Ho-Woo Nam, Zhao-shou Yang, Pan Li
    PLOS ONE.2023; 18(7): e0286430.     CrossRef
  • Past and present seroprevalence and disease burden estimates of Toxoplasma gondii infections in Germany: An appreciation of the role of serodiagnostics
    Frank Seeber
    International Journal of Medical Microbiology.2023; 313(6): 151592.     CrossRef
  • Seroprevalence of Toxoplasma gondii assayed using Rapid Diagnostic Tests among Residents in Three Counties Adjacent to The Demilitarized Zone, Korea
    Jeehi Jung, Jinyoung Lee, Yoon Kyung Chang, Seong Kyu Ahn, Seo Hye Park, Sung-Jong Hong, Jihoo Lee, Chom-Kyu Chong, Hye-Jin Ahn, Ho-Woo Nam, Tong-Soo Kim, Dongjae Kim
    The Korean Journal of Parasitology.2021; 59(1): 9.     CrossRef
  • Clusters of Toxoplasmosis in Ganghwa-gun, Cheorwon-gun, and Goseong-gun, Korea
    Jihye Yu, Woojin Kim, Yoon Kyung Chang, Tong-Soo Kim, Sung-Jong Hong, Hye-Jin Ahn, Ho-Woo Nam, Dongjae Kim
    The Korean Journal of Parasitology.2021; 59(3): 251.     CrossRef
  • Serological and molecular rapid diagnostic tests for Toxoplasma infection in humans and animals
    Amjad Hayat Khan, Rahmah Noordin
    European Journal of Clinical Microbiology & Infectious Diseases.2020; 39(1): 19.     CrossRef
  • Clusters of Toxoplasmosis in Gyodong-Myeon and Samsan-Myeon, Ganghwa-Gun, Korea
    Woojin Kim, Yoon Kyung Chang, Tong-Soo Kim, Sung-Jong Hong, Hye-Jin Ahn, Ho-Woo Nam, Dongjae Kim
    The Korean Journal of Parasitology.2020; 58(5): 493.     CrossRef
  • Identification of novel antigens for serum IgG diagnosis of human toxoplasmosis
    Juntao Luo, Jingyi Wan, Ziru Tang, Shuang Shen
    Experimental Parasitology.2019; 204: 107722.     CrossRef
  • Seroprevalence of Toxoplasmosis with ELISA and Rapid Diagnostic Test among Residents in Gyodong-do, Inchon city, Korea: A Four-Year Follow-up
    Yeong Hoon Kim, Ji hoo Lee, Seong kyu Ahn, Tong-Soo Kim, Sung-Jong Hong, Chom-Kyu Chong, Hye-Jin Ahn, Ho-Woo Nam
    The Korean Journal of Parasitology.2017; 55(3): 247.     CrossRef
  • Development of direct assays for Toxoplasma gondii and its use in genomic DNA sample
    Lívia M. Alves, Vinícius R. Rodovalho, Ana C.H. Castro, Márcia A.R. Freitas, Caroline M. Mota, Tiago W.P. Mineo, José R. Mineo, João M. Madurro, Ana G. Brito-Madurro
    Journal of Pharmaceutical and Biomedical Analysis.2017; 145: 838.     CrossRef
  • Seroprevalence of Toxoplasmosis Detected by RDT in Residents near the DMZ (demilitarized zone) of Cheorwon-gun, Gangwon-do, Korea
    Yeong Hoon Kim, Jihoo Lee, Young-Eun Kim, Seongkyu Ahn, Tong-Soo Kim, Sung-Jong Hong, Chom-Kyu Chong, Hye-Jin Ahn, Ho-Woo Nam
    The Korean Journal of Parasitology.2017; 55(4): 385.     CrossRef
  • Western Blot Detection of Human Anti-Chikungunya Virus Antibody with Recombinant Envelope 2 Protein
    Zhaoshou Yang, Jihoo Lee, Hye-Jin Ahn, Chom-Kyu Chong, Ronaldo F. Dias, Ho-Woo Nam
    The Korean Journal of Parasitology.2016; 54(2): 239.     CrossRef
  • Digested disorder, Quarterly intrinsic disorder digest (October-November-December, 2013)
    Shelly DeForte, Krishna D Reddy, Vladimir N Uversky
    Intrinsically Disordered Proteins.2015; 3(1): e984569.     CrossRef
  • Probability of Antibody Formation against Circumsporozoite Protein of Plasmodium vivax among Korean Malaria Patients
    Ho-Woo Nam, Kyoung Ju Song, Hye Jin Ahn, Zhaoshou Yang, Chom-Kyu Chong, Pyo Yun Cho, Seong Kyu Ahn, Tong-Soo Kim
    The Korean Journal of Parasitology.2014; 52(2): 143.     CrossRef
  • High Expression of Water-Soluble Recombinant Antigenic Domains ofToxoplasma gondii Secretory Organelles
    Zhaoshou Yang, Hye-Jin Ahn, Ho-Woo Nam
    The Korean Journal of Parasitology.2014; 52(4): 367.     CrossRef
  • 9,692 View
  • 106 Download
  • Crossref