Human Anti-Adenovirus IgG ELISA Kit

Name: Human Anti-Adenovirus IgG ELISA Kit
SKU: DEIA309
Rating: 5 (1 reviews)

Regulatory status: For research use only, not for use in diagnostic procedures.

COA

SDS

Datasheet

Summary

Sample

Serum, plasma

Species Reactivity

Human

Detection Method

iELISA

Intended Use

The ELISA test kit provides a in vitro assay for human antibodies of the IgG class against adenovirus in serum or plasma.

Contents of Kit

1. Microplate wells, coated with antigens: 12 microplate strips each containing 8 individual break-off wells in a frame, ready for use, 12 × 8. STRIPS.
2. Additional control / Calibrator 1, 200 RU/ml (IgG, human), ready for use, dark red, 1 × 2.0 ml. CAL 1.
3. Calibrator 2, 20 RU/ml (IgG, human), ready for use, red, 1 × 2.0 ml. CAL 2.
4. Additional control / Calibrator 3, 2 RU/ml (IgG, human), ready for use, light red, 1 × 2.0 ml. CAL 3.
5. Positive control (IgG, human), ready for use, blue, 1 × 2.0 ml. POS CONTROL.
6. Negative control (IgG, human), ready for use, green, 1 × 2.0 ml. NEG CONTROL.
7. Enzyme conjugate, peroxidase-labelled anti-human IgG (rabbit), ready for use, green, 1 × 12 ml. CONJUGATE.
8. Sample buffer, ready for use, light blue, 1 × 100 ml. SAMPLE BUFFER.
9. Wash buffer, 10× concentrate, colourless, 1 × 100 ml. WASH BUFFER 10×.
10. Chromogen/substrate solution, TMB/H₂O₂, ready for use, colourless, 1 × 12 ml. SUBSTRATE.
11. Stop solution, 0.5 M sulphuric acid, ready for use, colourless, 1 × 12 ml. STOP SOLUTION.
12. Test instruction --- 1 booklet
13. Protocol with target values --- 1 protocol

Storage

The test kit has to be stored at a temperature between +2°C to +8°C. Do not freeze. Unopened, all test kit components are stable until the indicated expiry date.
All reagents must be brought to room temperature (+18°C to +25°C) approx. 30 minutes before use. After initial opening, the reagents are stable until the indicated expiry date if stored at +2°C to +8°C and protected from contamination, unless stated othenwise below.

Precision

The reproducibility of the test was investigated by determining the intra- and inter-assay coefficients of variation using 3 sera. The intra-assay CVs are based on 20 determinations and the inter-assay CVs on 4 determinations performed in 6 different test runs.

Citations

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Convection Enhanced Delivery of the Oncolytic Adenovirus Delta24-RGD in Patients with Recurrent GBM: A Phase I Clinical Trial Including Correlative Studies

van Putten EHP, Kleijn A, van Beusechem VW, Noske D, Lamers CHJ, de Goede AL, Idema S, Hoefnagel D, Kloezeman JJ, Fueyo J, Lang FF, Teunissen CE, Vernhout RM, Bakker C, Gerritsen W, Curiel DT, Vulto A, Lamfers MLM, Dirven CMF

Clin Cancer Res2022 AprPubMed ID: 35176144Read Article

Applications: ELISA
Reactive species: Adenovirus
"Abstract: Purpose: Testing safety of Delta24-RGD (DNX-2401), an oncolytic adenovirus, locally delivered by convection enhanced delivery (CED) in tumor and surrounding brain of patients with recurrent glioblastoma. Patients and methods: Dose-escalation phase I study with 3+3 cohorts, dosing 107 to 1 × 1011 viral particles (vp) in 20 patients. Besides clinical parameters, adverse events, and radiologic findings, blood, cerebrospinal fluid (CSF), brain interstitial fluid, and excreta were sampled over time and analyzed for presence of immune response, viral replication, distribution, and shedding. Results: Of 20 enrolled patients, 19 received the oncolytic adenovirus Delta24-RGD, which was found to be safe and feasible. Four patients demonstrated tumor response on MRI, one with complete regression and still alive after 8 years. Most serious adverse events were attributed to increased intracranial pressure caused by either an inflammatory reaction responding to steroid treatment or viral meningitis being transient and self-limiting. Often viral DNA concentrations in CSF increased over time, peaking after 2 to 4 weeks and remaining up to 3 months. Concomitantly Th1- and Th2-associated cytokine levels and numbers of CD3+ T and natural killer cells increased. Posttreatment tumor specimens revealed increased numbers of macrophages and CD4+ and CD8+ T cells. No evidence of viral shedding in excreta was observed. Conclusions: CED of Delta24-RGD not only in the tumor but also in surrounding brain is safe, induces a local inflammatory reaction, and shows promising clinical responses."
Article snippet: Serum and cerebrospinal fluid immunoglobulin antibodies against adenovirus were determined quantitatively by adenovirus IgG and adenovirus IgM ELISA kits (Creative Diagnostics) according to manufacturer’s instructions. For both IgG and IgM assays, samples obtained before virus treatment and at 4 weeks after were thawed and duplicate serial dilutions up to 1:2,000 were prepared followed by testing against the provided standard curves of the kits. The absorbance was read on a VERSAmax tunable microplate reader (****). Results are expressed in arbitrary units (U/mL).

Levels are expressed in units per mL serum. Anti-adenovirus IgG antibodies found in the CSF of patients prior to (white bars) and 4 weeks after (black bars) Delta24-RGD treatment are shown in B. (Valenzuela-Leon PC, <i>et al</i>, 2022)

Figure 1. Serum-specific anti-adenovirus IgG antibodies were determined in all patients prior to (white bars) and 4 weeks after (black bars) Delta24-RGD treatment.

Background

Adenoviruses are non-enveloped DNA viruses that can infect a wide range of vertebrate hosts, and the clinical manifestations of human infections vary from mild respiratory infections to life-threatening multi-organ diseases. Human adenoviruses are classified into different serotypes based on whether they are cross-neutralized by antibodies, and more than 50 disease-causing serotypes have been identified, divided into 6 subgroups from A to F. Of these, human adenovirus serotypes 1, 2, and 5 primarily cause mild respiratory illness, while human adenovirus serotypes 4 and 7 may cause more severe pneumonia. Due to their high infectivity, adenovirus is mainly used as a vector for gene transfer. Adenoviruses have many advantages as vectors, firstly, the viral genome has a clear structure and is easy to manipulate for insertion of exogenous genes. Secondly, adenovirus has a very broad host cell tropism and can effectively infect cells. In addition, recombinant genomes are stable for continuous transmission and can be produced rapidly and on a large scale. In addition to these advantages, when recombinant adenovirus vaccines are used by the mucosal route, high transduction efficiencies are expected because mucosal infection is an inherent property of adenoviruses

The adenoviral genome has been genetically engineered for safe and effective use of adenoviral vectors. The first-generation of adenoviral vectors knocked out E1/E3, allowing for transgene insertion, replication-defective characterization, and improved immunogenicity. However, this generation of vector genomes has homology to the E1 portion of the insertional packaging cell line HEK293, and therefore it is likely that replication-competent adenovirus (RCA) will emerge. The second-generation adenoviral vectors further improved transgenic capacity by additional deletion of the E2/E4 locus, reducing the likelihood of RCA formation. However, the overall yield was lower than the first-generation adenoviral vectors due to the reduced replication capacity of the production cell lines. The third generation of adenoviral vectors deletes almost all genomic sequences except those necessary for packaging, and is therefore also known as helper gene-dependent adenovirus vectors. It can also insert multiple transgene expression cassettes and has a high capacity, but it is more difficult to manufacture, less immunogenic than previous generations, and still has the potential to be contaminated with helper viruses.

Adenoviral genome and recombinant vector modification Figure 1. Schematic representation of the adenovirus genome and adenovirus-based vectors
(Source: Chang J. 2021)

The versatility and efficacy of adenoviral vectors have made them a useful platform for vaccines against new pathogens or vaccines in need of improvement, and with continued research and improvement, adenoviral vectors will become an indispensable tool in vaccine development.

Alternative Names

Human Anti-AdV
Anti-HAd

References

1. Chang J. Adenovirus Vectors: Excellent Tools for Vaccine Development. Immune Netw. 2021 Feb 15;21(1):e6.
2. Gallardo J, et al. Adenovirus Structure: What Is New? Int J Mol Sci. 2021 May 15;22(10):5240.

Documents

pdf COA

pdf SDS

pdf Datasheet

Reagents for Adenovirus Assay Development

QA & Reviews

Customer Reviews

References

Adenovirus Infection and Transplantation

Transplantation

Authors: Al-Heeti OM, Cathro HP, Ison MG.

Abstract

Adenoviruses result in a wide array of clinical presentations, including primarily respiratory, gastrointestinal, genitourinary, or systemic infections. Although adenovirus causes mild disease limited to a single organ system in immunocompetent individuals, severe and life-threatening infections do rarely occur. Disseminated disease and severe localized disease resulting in significant morbidity and mortality have been well described in the immunocompromised populations. Although asymptomatic viremia, respiratory tract, and gastrointestinal infections are the most common disease in most transplant patients, renal transplant patients more commonly experience urinary tract infections, including hemorrhagic cystitis or nephritis. Diagnosis requires astute clinical awareness of the patient's clinical presentation that would be compatible with adenovirus combined with cultures, molecular testing, polymerase chain reaction, and tissue sampling. There is no Food and Drug Administration-approved treatment for adenovirus; however, several studies have evaluated therapeutic options including cidofovir, brincidofovir, and immunotherapy. This article will summarize our current understanding of adenovirus in the transplant population

Cytokine Responses to Adenovirus and Adenovirus Vectors

Viruses

Authors: Atasheva S, Shayakhmetov DM.

Abstract

The expression of cytokines and chemokines in response to adenovirus infection is tightly regulated by the innate immune system. Cytokine-mediated toxicity and cytokine storm are known clinical phenomena observed following naturally disseminated adenovirus infection in immunocompromised hosts as well as when extremely high doses of adenovirus vectors are injected intravenously. This dose-dependent, cytokine-mediated toxicity compromises the safety of adenovirus-based vectors and represents a critical problem, limiting their utility for gene therapy applications and the therapy of disseminated cancer, where intravenous injection of adenovirus vectors may provide therapeutic benefits. The mechanisms triggering severe cytokine response are not sufficiently understood, prompting efforts to further investigate this phenomenon, especially in clinically relevant settings. In this review, we summarize the current knowledge on cytokine and chemokine activation in response to adenovirus- and adenovirus-based vectors and discuss the underlying mechanisms that may trigger acute cytokine storm syndrome. First, we review profiles of cytokines and chemokines that are activated in response to adenovirus infection initiated via different routes. Second, we discuss the molecular mechanisms that lead to cytokine and chemokine transcriptional activation. We further highlight how immune cell types in different organs contribute to synthesis and systemic release of cytokines and chemokines in response to adenovirus sensing. Finally, we review host factors that can limit cytokine and chemokine expression and discuss currently available and potential future interventional approaches that allow for the mitigation of the severity of the cytokine storm syndrome. Effective cytokine-targeted interventional approaches may improve the safety of systemic adenovirus delivery and thus broaden the potential clinical utility of adenovirus-based therapeutic vectors.