Rapid antibody quantification and generation of whole proteome antibody response profiles using LIPS (luciferase immunoprecipitation systems)

Burbelo, Peter D.; Ching, Kathryn H.; Mattson, Thomas L.; Light, Jason S.; Bishop, Lisa R.; Kovacs, Joseph A.

doi:10.1016/j.bbrc.2006.11.140

Cited by 61 publications

(77 citation statements)

References 14 publications

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“…Briefly, animal sera were processed in a 96-well format at room temperature as previously described (6,8,9). Serum samples were first diluted 1:10 in assay buffer A (50 mM Tris, pH 7.5, 100 mM NaCl, 5 mM MgCl 2 , 1% Triton X-100) using a 96-well polypropylene microtiter plate.…”

Section: Methodsmentioning

confidence: 99%

“…Similarly, although the demonstration of specific adaptive immune responses against viral structural and nonstructural proteins cannot in itself prove a causal relationship to disease, it does provide definitive evidence of host infection (2,7). Here, we describe the usefulness of a sensitive serological assay (7)(8)(9)(10)(11) that can be quickly established for new pathogens to screen different animal species for evidence of virus infection and thereby for identification of a novel virus's natural reservoir and host range. Our serology data indicated the presence of CHV-like viruses in horses and led to the genetic characterization of eight novel and genetically diverse nonprimate hepaciviruses (NPHVs).…”

mentioning

confidence: 99%

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Serology-Enabled Discovery of Genetically Diverse Hepaciviruses in a New Host

Burbelo

Dubovi

Simmonds

et al. 2012

J Virol

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215

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Genetic and biological characterization of new hepaciviruses infecting animals contributes to our understanding of the ultimate origins of hepatitis C virus (HCV) infection in humans and dramatically enhances our ability to study its pathogenesis using tractable animal models. Animal homologs of HCV include a recently discovered canine hepacivirus (CHV) and GB virus B (GBV-B), both viruses with largely undetermined natural host ranges. Here we used a versatile serology-based approach to determine the natural host of the only known nonprimate hepacivirus (NPHV), CHV, which is also the closest phylogenetic relative of HCV. Recombinant protein expressed from the helicase domain of CHV NS3 was used as antigen in the luciferase immunoprecipitation system (LIPS) assay to screen several nonprimate animal species. Thirty-six samples from 103 horses were immunoreactive, and viral genomic RNA was present in 8 of the 36 seropositive animals and none of the seronegative animals. Complete genome sequences of these 8 genetically diverse NPHVs showed 14% (range, 6.4% to 17.2%) nucleotide sequence divergence, with most changes occurring at synonymous sites. RNA secondary structure prediction of the 383-base 5′ untranslated region of NPHV was refined and extended through mapping of polymorphic sites to unpaired regions or (semi)covariant pairings. Similar approaches were adopted to delineate extensive RNA secondary structures in the coding region of the genome, predicted to form 27 regularly spaced, thermodynamically stable stem-loops. Together, these findings suggest a promising new nonprimate animal model and provide a database that will aid creation of functional NPHV cDNA clones and other novel tools for hepacivirus studies.

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Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

Serology-Enabled Discovery of Genetically Diverse Hepaciviruses in a New Host

Burbelo

Dubovi

Simmonds

et al. 2012

J Virol

Self Cite

215

343

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“…A method known as luciferase immunoprecipitation systems (LIPS) quantitatively measures antibodies to a wide range of infectious agents, [16][17][18][19] as well as to a variety of human autoantigens. [20][21][22] LIPS is a liquid phase immunoassay that uses antigens directly tagged with Renilla luciferase, which can sensitively and quantitatively detect antibodies.…”

Section: Functional Testing Proved That Autoantibodies Directed Againmentioning

confidence: 99%

Anti-cytokine autoantibodies are associated with opportunistic infection in patients with thymic neoplasia

Burbelo

Browne

Sampaio

et al. 2010

Blood

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Patients with thymic malignancy have high rates of autoimmunity leading to a variety of autoimmune diseases, most commonly myasthenia gravis caused by anti-acetylcholine receptor autoantibodies. High rates of autoantibodies to cytokines have also been described, although prevalence, spectrum, and functionality of these anti-cytokine autoantibodies are poorly defined. To better understand the presence and function of anti-cytokine autoantibodies, we created a luciferase immunoprecipitation system panel to search for autoantibodies against 39 different cytokines and examined plasma from controls (n ‫؍‬ 30) and patients with thymic neoplasia (n ‫؍‬ 17). In this screen, our patients showed statistically elevated, but highly heterogeneous immunoreactivity against 16 of the 39 cytokines. Some patients showed autoantibodies to multiple cytokines.

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“…LIPS can distinguish L. loa infection from other filarial infections. Sera from patients with filarial infections often have broad filarial antigen cross-reactivity (3,4). Because of the close phylogenetic relationships among the filarial species (Fig.…”

Section: Lips Detection Of Anti-llsxp-1 Antibodies For Diagnosis Of Lmentioning

confidence: 99%

“…We have recently described a highly sensitive immunoprecipitation technology, designated the luciferase immunoprecipitation system (LIPS), that utilizes mammalian cell-produced, recombinant fusion protein antigens for efficiently evaluating antibody responses (4)(5)(6). In the present study, LIPS technology was used to develop an assay for L. loa infection that is rapid, sensitive, specific, and high throughput.…”

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confidence: 99%

Rapid, Novel, Specific, High-Throughput Assay for Diagnosis of Loa loa Infection

et al. 2008

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The ability to diagnose Loa loa infection readily and accurately remains a demanding task. Among the available diagnostic methods, many are impractical for point-of-care field testing. To investigate whether luciferase immunoprecipitation systems (LIPS) can be used for rapid and specific diagnosis of L. loa infection, a LIPS assay was developed based on immunoglobulin G (IgG) and IgG4 subclass antibodies to a recombinant L. loa SXP-1 (designated LlSXP-1) antigen and tested with sera from healthy controls or patients with proven infection with L. loa, Mansonella perstans, Onchocerca volvulus, Strongyloides stercoralis, or Wuchereria bancrofti. A LIPS test measuring IgG antibody against LlSXP-1 readily differentiated L. loa-infected from uninfected patients and demonstrated markedly improved sensitivity and specificity compared with an LlSXP-1 IgG4-based enzyme-linked immunosorbent assay (67% sensitivity and 99% specificity). No significant immunoreactivity was observed with S. stercoralisinfected sera, but a small number of patients infected with O. volvulus, M. perstans, or W. bancrofti showed positive immunoreactivity. Measuring anti-IgG4-specific antibodies to LlSXP-1 showed a significant correlation (r ϳ 0.85; P < 0.00001) with the anti-IgG results but showed no advantage over measuring the total IgG response alone. In contrast, a rapid LIPS format (called QLIPS) in which the tests are performed in less than 15 minutes under nonequilibrium conditions significantly improved the specificity for cross-reactive O. volvulus patient sera (100% sensitivity and 100% specificity). These results suggest that LIPS (and the even more rapid test QLIPS) represents a major advance in the ability to diagnose L. loa infection and may have future applications for point-of-care diagnostics.The development of rapid diagnostic assays for onchocerciasis (22) and lymphatic filariasis (20) has not only simplified the care of individual patients with these filarial infections but also allowed accurate and cost-effective geographic mapping for the purpose of mass chemotherapy in areas of endemicity (21) and for use in the certification process of elimination (15). Nevertheless, the coendemicity of loiasis with these and other filarial infections of humans remains an issue because of species-specific differences in the responses to available antifilarial therapies. This has been of particular concern in the setting of both the African Program for Onchocerciasis Control (APOC) and the Global Program for the Elimination of Lymphatic Filariasis (GPELF), where mass drug administration has ground to a halt in certain areas of Africa because of deaths related to ivermectin administered as part of mass treatment programs for onchocerciasis control (3,12).Although the definitive diagnosis of Loa loa infection can be made morphologically by identifying microfilariae in the blood or, rarely, after surgical removal of the adult worm (typically from its subconjunctival location), a proportion of infected individuals are amicrofilaremic (9, 13). PC...

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Rapid antibody quantification and generation of whole proteome antibody response profiles using LIPS (luciferase immunoprecipitation systems)

Cited by 61 publications

References 14 publications

Serology-Enabled Discovery of Genetically Diverse Hepaciviruses in a New Host

Serology-Enabled Discovery of Genetically Diverse Hepaciviruses in a New Host

Anti-cytokine autoantibodies are associated with opportunistic infection in patients with thymic neoplasia

Rapid, Novel, Specific, High-Throughput Assay for Diagnosis of Loa loa Infection

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