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.
Highlights
P24 protein is derived from the Gag protein of Human immunodeficiency virus type 1 (HIV-1) and plays an important role in viral core assembly and maturation [1, 2]
The purified protein of interest was confirmed by Enzyme linked immunosorbent assay (ELISA) analysis of the protein using commercially available monoclonal antibody against p24 antigen (Figure 1(b))
With the introduction of rapid HIV antibody test kits, HIV screening has become more decentralized with more tests done on an individual rather than batch [21]
Summary
P24 protein is derived from the Gag protein of HIV-1 and plays an important role in viral core assembly and maturation [1, 2]. HIV-1 RNA, anti-HIV antibodies, and p24 antigen are viral markers which have been used as a target antigen for early detection of HIV-1 infection [3, 4]. Over the past two decades, HIV immunoassays have gone through first-generation (using viral lysate for IgG antibody detection), second-generation (using recombinant antigens for IgG antibody detection), third-generation (IgM and IgG antibodies detection), and fourth-generation (antibody and p24 antigen detection) immunoassay. These test kits have helped to shorten the “window period” and provide an early diagnosis for suspected HIV infected samples as compared with the third-generation immunoassays. MAbs were screened against recombinant p24 protein and its application in GICA for HIV-1 detection was explored
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
