The use of epitope arrays in immunodiagnosis of infectious disease: Hepatitis C virus, a case study

Siman-Tov, Dror D.; Zemel, Romy; Kaspa, Ran Tur; Gershoni, Jonathan M.

doi:10.1016/j.ab.2012.09.025

Cited by 9 publications

(6 citation statements)

References 45 publications

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“…As requested by CDC, the second assay is imperative to circumvent the false-positive bias of ELISA and confirm the diagnostic result, , such as recombinant immunoblot assay (RIBA). It is also a serologic immunoassay for IgG antibody but employs multiple individual antigens to report a reaction pattern, thus inherently providing a higher level of specificity than ELISA. , However, its routine utilization might be hampered by high cost (at least 2-fold that of ELISA), low sensitivity, and long assay time (6–7 h) . Most importantly, the segmented clinical pathway itself might be a major reason to hinder the effective delivery of diagnosis by adding diagnostic complexity, delaying turnaround time and increasing office visits.…”

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confidence: 99%

Multiplex Microfluidic Paper-based Immunoassay for the Diagnosis of Hepatitis C Virus Infection

et al. 2014

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Hepatitis C virus (HCV) infection is a serious and rising global healthcare problem. One critical challenge to tackle this disease is the lack of adequate diagnosis. Here, we develop a multiplex microfluidic paper-based immunoassay, as a novel diagnostic approach, to detect human IgG antibody against HCV (anti-HCV). The paper substrate, highly flammable nitrocellulose (NC), is patterned under ambient temperature by craft punch patterning (CPP) to generate multiple test zones. On the basis of superior merits of patterned paper, this new diagnostic approach demonstrates the key novelty to unprecedentedly combine segmented diagnostic assays into a single multiplex test. The generated diagnostic results are not only informative but can be rapidly and cost-effectively delivered. It would significantly transform the clinical pathway for unwitting individuals with HCV infection. This work highlights the promising role of microfluidic paper-based immunoassays in tackling the diagnostic challenge for the HCV pandemic as well as other diseases.

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confidence: 99%

Multiplex Microfluidic Paper-based Immunoassay for the Diagnosis of Hepatitis C Virus Infection

et al. 2014

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“…The murine anti-gp120 mAb CG10, which stringently recognizes the CD4i conformation of gp120 [ 14 , 17 , 38 , 39 ], the murine anti-gp120 mAbs LG4 (targets a conserved epitope at the carboxy-terminus of gp120), 9G3 and 1B6 [ 85 ], the murine anti-CD4 mAb CG9 [ 14 ], the murine anti-M13 Y2D mAb [ 86 ] and the rabbit polyclonal anti-M13 serum were produced at Tel Aviv University. The human anti-gp120 mAb N12-i15, which stringently recognizes the CD4i conformation of gp120, was produced at the Institute of Human Virology, University of Maryland School of Medicine, Baltimore, MA, USA [ 15 , 25 , 40 ].…”

Section: Methodsmentioning

confidence: 99%

Allosteric induction of the CD4-bound conformation of HIV-1 Gp120

et al. 2013

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BackgroundHIV-1 infection of target cells is mediated via the binding of the viral envelope protein, gp120, to the cell surface receptor CD4. This interaction leads to conformational rearrangements in gp120 forming or revealing CD4 induced (CD4i) epitopes which are critical for the subsequent recognition of the co-receptor required for viral entry. The CD4-bound state of gp120 has been considered a potential immunogen for HIV-1 vaccine development. Here we report on an alternative means to induce gp120 into the CD4i conformation.ResultsCombinatorial phage display peptide libraries were screened against HIV-1 gp120 and short (14aa) peptides were selected that bind the viral envelope and allosterically induce the CD4i conformation. The lead peptide was subsequently systematically optimized for higher affinity as well as more efficient inductive activity. The peptide:gp120 complex was scrutinized with a panel of neutralizing anti-gp120 monoclonal antibodies and CD4 itself, illustrating that peptide binding does not interfere with or obscure the CD4 binding site.ConclusionsTwo surfaces of gp120 are considered targets for the development of cross neutralizing antibodies against HIV-1; the CD4 binding site and CD4i epitopes. By implementing novel peptides that allosterically induce the CD4i epitopes we have generated a viral envelope that presents both of these surfaces simultaneously.

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“…The human mAbs 80R (Sui et al, 2004) and 11A (Sui et al, 2008), which target the RBD of the SARS CoV, and the human ACE2 protein were produced at the Dana-Farber Cancer Institute, Boston, MA. Polyclonal rabbit anti-M13 sera and the recombinant pVIII-specific GIL-Ab (Kaplan and Gershoni, 2012) as well as the anti-m13 mAb Y2D (Siman-Tov et al, 2013) were produced at Tel Aviv University.…”

Section: Monoclonal Antibodies and Proteinsmentioning

confidence: 99%

Reconstitution of the receptor-binding motif of the SARS coronavirus

Freund

Roitburd-Berman

Sui

et al. 2015

Protein Engineering, Design and Selection

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The severe acute respiratory syndrome (SARS) coronavirus (CoV) identified in 2003 has infected ∼8000 people worldwide, killing nearly 10% of them. The infection of target cells by the SARS CoV is mediated through the interaction of the viral Spike (S) protein (1255 amino acids) and its cellular receptor, angiotensin-converting enzyme 2 (ACE2). The SARS CoV receptor-binding domain (amino acids N318-T509 of S protein) harbors an extended excursion along its periphery that contacts ACE2 and is designated the receptor-binding motif (RBM, amino acids S432-T486). In addition, the RBM is a major antigenic determinant, able to elicit production of neutralizing antibodies. Hence, the role of the RBM is a bi-functional bioactive surface that can be demonstrated by antibodies such as the neutralizing human anti-SARS monoclonal antibody (mAb) 80R which targets the RBM and competes with the ACE2 receptor for binding. Here, we employ phage-display peptide-libraries to reconstitute a functional RBM. This is achieved by generating a vast collection of candidate RBM peptides that present a diversity of conformations. Screening such 'Conformer Libraries' with corresponding ligands has produced short RBM constructs (ca. 40 amino acids) that can bind both the ACE2 receptor and the neutralizing mAb 80R.

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The use of epitope arrays in immunodiagnosis of infectious disease: Hepatitis C virus, a case study

Cited by 9 publications

References 45 publications

Multiplex Microfluidic Paper-based Immunoassay for the Diagnosis of Hepatitis C Virus Infection

Multiplex Microfluidic Paper-based Immunoassay for the Diagnosis of Hepatitis C Virus Infection

Allosteric induction of the CD4-bound conformation of HIV-1 Gp120

Reconstitution of the receptor-binding motif of the SARS coronavirus

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