A Unique Monoclonal Antibody That Recognizes Mature p17 of HIV‐1 but Not Its Precursor

Saitoh, Atsushi; Tanaka, Naoto; Nakata, Atsuo; Ikuta, Kazuyoshi; Shinagawa, Hideo

doi:10.1111/j.1348-0421.1992.tb01647.x

Cited by 24 publications

(17 citation statements)

References 17 publications

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“…The results of SDS-PAGE of the cells with pTG171 and pTG172 were indistinguishable (Fig. 2, lanes 4 and 5), however, unprocessed products as well as p17 were observed in the cells with pTG171, while unprocessed product was not observed in those with pTG172 by immunoblotting as reported previously (39). To ensure exclusive production of each of the Gag proteins, we decided to use pTG172 and pTG120 for the preparation of p17 and p24, respectively.…”

Section: Resultsmentioning

confidence: 83%

“…In the cells with pTG 171, unprocessed products were not identified by SDS-PAGE (Fig. 2, lane 4) but were observed by immunoblotting as reported previously (39). To increase the expression of protease for efficient processing of the Gag proteins, the pol gene was fused in-frame to the gag gene to be expressed without frameshifting in pTG592, pTG120, and pTG172.…”

Section: Discussionmentioning

confidence: 88%

“…The Gag-Pol proteins, which were synthesized through frameshifting as the minor products, containing the entire protease moiety, had the protease activity and could process the Gag proteins synthesized as major products and themselves in E. coli (39,43). However, the protease synthesized via frameshifting could not process the Gag proteins completely.…”

Section: Discussionmentioning

confidence: 99%

“…We and others have reported that the gag-to-pol frameshifting occurred during the translation of the gag gene in E. coli, as in the virus-infected cells (38,39,43). The Gag-Pol proteins, which were synthesized through frameshifting as the minor products, containing the entire protease moiety, had the protease activity and could process the Gag proteins synthesized as major products and themselves in E. coli (39,43).…”

Section: Discussionmentioning

confidence: 99%

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Overproduction, Purification, and Diagnostic Use of the Recombinant HIV‐1 Gag Proteins, the Precursor Protein p55 and the Processed Products p17, p24, and p15

Saito

Morimoto

Ohara

et al. 1995

Microbiology and Immunology

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HIV-1 Gag protein precursor p55, and its processed products, p17, p24, and p15 were overproduced in Escherichia coli and purified to near homogeneity. To study the antigenic properties and the potentiality as the diagnostic and prognostic reagents, varying amounts of the purified Gag proteins were dotted onto the polyvinylidene difluoride membrane and reacted with 40 sera of HIV-1-infected individuals (35 AC, 1 ARC, and 4 AIDS patients) and 10 sera of normal healthy donors. p55 reacted with 40 (100%) sera of HIV-1 carriers, while p17, p24, and p15 reacted with 37 (92.5%), 35 (87.5%) and 34 (85%) of the 40 sera of HIV-1 carriers, respectively. On the whole, the reaction of p55 was especially strong and that of p15 was the weakest. p55 showed the strongest reaction among the four Gag proteins with all specimens, and it showed a positive reaction with a carrier serum with which none of the processed Gag proteins showed a positive reaction. Therefore, p55 is the most useful antigen among the four Gag proteins for detection of the Gag antibodies and may even be one of the most useful antigens for the diagnosis of HIV-1 infection.

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Section: Resultsmentioning

confidence: 83%

Section: Discussionmentioning

confidence: 88%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

See 2 more Smart Citations

Overproduction, Purification, and Diagnostic Use of the Recombinant HIV‐1 Gag Proteins, the Precursor Protein p55 and the Processed Products p17, p24, and p15

Saito

Morimoto

Ohara

et al. 1995

Microbiology and Immunology

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“…Recombinant HIV-1 p17 (rp17) and p24 (rp24) antigens were produced in Escherichia coli transformed with expression plasmids carrying the corresponding cDNAs and were purified as described previously (28,31). The recombinant proviral clone used was pNL4-3 (1), which contained DNA from HIV-1 isolates NY5 (GenBank accession no.…”

Section: Methodsmentioning

confidence: 99%

Earlier Detection of Human Immunodeficiency Virus Type 1 p24 Antigen and Immunoglobulin G and M Antibodies to p17 Antigen in Seroconversion Serum Panels by Immune Complex Transfer Enzyme Immunoassays

Hashida

Ishikawa

Hashinaka

et al. 2000

Clin Diagn Lab Immunol

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For earlier diagnosis of human immunodeficiency virus type 1 (HIV-1) infection, the sensitivities of immune complex transfer enzyme immunoassays for HIV-1 p24 antigen and antibody immunoglobulin G (IgG) to HIV-1 p17 antigen were improved approximately 25-and 90-fold, respectively, over those of the previous immunoassays by performing solid-phase immunoreactions with shaking and increasing the serum sample volumes, and immune complex transfer enzyme immunoassay of antibody IgM to p17 antigen was also performed in the same way as the improved immunoassay of antibody IgG to p17 antigen. By the improved immunoassays, p24 antigen and antibody IgG to p17 antigen were detected earlier in 32 and 53%, respectively, of the HIV-1 seroconversion serum panels tested than before the improvements, and p24 antigen was detected as early as or earlier than HIV-1 RNA by reverse transcriptase-PCR (RT-PCR) in all of the panels tested. In 4 panels out of 19 tested, antibody IgG to p17 antigen or both antibodies IgG and IgM to p17 antigen were detected earlier than p24 antigen and RNA, although the antibody levels declined slightly before their steep increases usually observed after p24 antigen and RNA. Thus, the window period in diagnosis of HIV-1 infection can be shortened by detection of p24 antigen with the improved immunoassay as much as by detection of RNA with RT-PCR and, in some cases, more by detection of antibodies IgG and IgM to p17 antigen with the improved immunoassays than by detections of p24 antigen with the improved immunoassay and RNA with RT-PCR.The positive rates of immunoglobulin G (IgG) antibodies to human immunodeficiency virus type 1 (HIV-1) antigens in serum samples from HIV-1-infected subjects have been reported by the conventional enzyme-linked immunosorbent assay (ELISA) and Western blotting, and it has been generally accepted that the positive rates are high with gp160, gp41 and reverse transcriptase (RT), or p66 (a subunit of RT) as antigens (98 to 100% in asymptomatic carriers, 86 to 100% in patients with AIDS-related complex [ARC], and 77 to 100% in patients with AIDS) but low with gag proteins as antigens (63 to 92% in asymptomatic carriers, 50 to 97% in patients with ARC, and 21 to 77% in patients with AIDS using p24 as antigen and 41% in asymptomatic carriers, 30% in patients with ARC, and 14% in patients with AIDS using p17 as antigen) (2,4,6,8). In seroconversion serum panels, the earliest positive band seen by Western blotting has been observed with p24 in some cases (26,29,34) and with gp160 in other cases (30, 32, 35) but not with p17 (13).Recently, ultrasensitive enzyme immunoassays (EIAs) (immune complex transfer EIAs) of antibody IgGs to HIV-1 antigens have been developed using recombinant RT, p24, and p17 antigens (10,12,18). Antibody IgGs were allowed to react simultaneously with 2,4-dinitrophenyl antigens and antigenenzyme conjugates. The immune complexes of the three components formed were trapped on (anti-2,4-dinitrophenyl group) IgG-coated solid-phase (first solid phase) and, after wash...

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Earlier diagnosis of HIV-1 infection by simultaneous detection of p24 antigen and antibody IgGs to p17 and reverse transcriptase in serum with enzyme immunoassay

Hashida¹,

Hashinaka²,

Nishikata³

et al. 1996

J. Clin. Lab. Anal.

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A Unique Monoclonal Antibody That Recognizes Mature p17 of HIV‐1 but Not Its Precursor

Cited by 24 publications

References 17 publications

Overproduction, Purification, and Diagnostic Use of the Recombinant HIV‐1 Gag Proteins, the Precursor Protein p55 and the Processed Products p17, p24, and p15

Overproduction, Purification, and Diagnostic Use of the Recombinant HIV‐1 Gag Proteins, the Precursor Protein p55 and the Processed Products p17, p24, and p15

Earlier Detection of Human Immunodeficiency Virus Type 1 p24 Antigen and Immunoglobulin G and M Antibodies to p17 Antigen in Seroconversion Serum Panels by Immune Complex Transfer Enzyme Immunoassays

Earlier diagnosis of HIV-1 infection by simultaneous detection of p24 antigen and antibody IgGs to p17 and reverse transcriptase in serum with enzyme immunoassay

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