title: Development of Monoclonal Antibodies against HIV-1 p24 Protein and Its Application in Colloidal Gold Immunochromatographic Assay for HIV-1 Detection.
abstract: Human immunodeficiency virus type 1 (HIV-1) p24 protein is the most abundant viral protein of HIV-1. This protein is secreted in blood serum at high levels during the early stages of HIV-1 infection, making it a biomarker for early diagnosis. In this study, a colloidal gold immunochromatographic assay (GICA) was established for detecting p24 protein using mouse monoclonal antibodies (mAbs). The HIV-1 p24 protein was expressed in E. coli strain BL21 and the purified protein was used to immunize mice. Stable hybridoma cell lines secreting anti-p24 monoclonal antibodies were obtained after ELISA screening and subcloning by limiting dilution. 34 different capture and labeling mAb pairs were selected by a novel antibody-capture indirect sandwich ELISA and then applied in GICA to detect p24 protein. The GICA method has a limit of detection (LOD) of 25 pg/mL and could detect p24 protein in all 10 positive samples obtained from the National Reference of HIV-1 p24 antigen. Out of 153 negative samples tested, 3 false positives results were obtained. The overall specificity of this test was 98.03%. The good sensitivity and specificity of this method make it a suitable alternative to provide a more convenient and efficient tool for early diagnosis of HIV infection.
authors.affiliation: School of Bioscience and Bioengineering, South China University of Technology, Guangzhou 510006, China.
authors.forename: Yi
authors.initials: Y
authors.lastname: Ma
authors.affiliation: School of Bioscience and Bioengineering, South China University of Technology, Guangzhou 510006, China.
authors.forename: Chao
authors.initials: C
authors.lastname: Ni
authors.affiliation: School of Bioscience and Bioengineering, South China University of Technology, Guangzhou 510006, China.
authors.forename: Emmanuel E
authors.initials: EE
authors.lastname: Dzakah
authors.affiliation: School of Bioscience and Bioengineering, South China University of Technology, Guangzhou 510006, China.
authors.forename: Haiying
authors.initials: H
authors.lastname: Wang
authors.affiliation: National Engineering Laboratory of Rapid Diagnostic Tests, Guangzhou Wondfo Biotech Co., Ltd., Guangzhou 510663, China.
authors.forename: Keren
authors.initials: K
authors.lastname: Kang
authors.affiliation: National Engineering Laboratory of Rapid Diagnostic Tests, Guangzhou Wondfo Biotech Co., Ltd., Guangzhou 510663, China.
authors.forename: Shixing
authors.initials: S
authors.lastname: Tang
authors.affiliation: National Engineering Laboratory of Rapid Diagnostic Tests, Guangzhou Wondfo Biotech Co., Ltd., Guangzhou 510663, China.
authors.forename: Jihua
authors.initials: J
authors.lastname: Wang
authors.affiliation: School of Bioscience and Bioengineering, South China University of Technology, Guangzhou 510006, China.
authors.forename: Jufang
authors.initials: J
authors.lastname: Wang
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D001688:Biological Products; D002846:Chromatography, Affinity / Q000379:methods*; D004926:Escherichia coli; D017979:Gold Colloid / Q000737:chemistry*; D016655:HIV Core Protein p24 / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D015658:HIV Infections / Q000175:diagnosis / Q000821:virology; D015497:HIV-1 / Q000276:immunology / Q000302:isolation & purification*; D006801:Humans; D057230:Limit of Detection; D051379:Mice; D008807:Mice, Inbred BALB C; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism
chemical_list: D000911:Antibodies, Monoclonal; D001688:Biological Products; D017979:Gold Colloid; D016655:HIV Core Protein p24; D011994:Recombinant Proteins; C514799:p24 protein, Human Immunodeficiency Virus Type 1