Posted in Antibodies, Clia Kits, Culture Cells, DNA, Test Kits
A cell-based ELISA as surrogate of virus neutralization assay for RBD SARS-CoV-2 specific antibodies
Lawrence- posted on
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SARS-CoV-2, the cause of the COVID-19 pandemic, has provoked a global crisis and death of millions of people. Several serological assays to determine the quality of the immune response against SARS-CoV-2 and the efficacy of vaccines have been developed, among them the gold standard conventional virus neutralization assays.
However, Bio Med Frontiers these tests are time-consuming, require biosafety level 3 (BSL3), and are low throughput and expensive.
This has motivated the development of alternative methods, including molecular inhibition assays.
Herein, we present a safe cell-based ELISA-virus neutralization test (cbE-VNT) as a surrogate for the conventional viral neutralization assays that detect the inhibition of SARS-CoV-2 RBD binding to ACE2-bearing cells independently of species.
Our test shows a very good correlation with the conventional and molecular neutralization assays and achieves 100% specificity and 95% sensitivity. cbE-VNT is cost-effective, fast and enables a large-scale serological evaluation that can be performed in a BSL2 laboratory, allowing its use in pre-clinical and clinical investigations.
| Rabbit Anti BSA | |||
| MyBiosource | |||
| Rabbit Anti BSA | |||
| MyBiosource | |||
| Rabbit Anti BSA | |||
| MyBiosource | |||
| Rabbit Anti BSA | |||
| MyBiosource | |||
| Rabbit Anti BSA | |||
| MyBiosource | |||
Comparative performance of ELISA and dot blot assay for TSH-receptor antibody detection in Graves’ disease.
Graves’ disease (GD) is an autoimmune disease, and it accounts for major cases of hyperthyroidism. Antibody against thyroid-stimulating hormone receptor/TSHR (TRAb) is responsible for hyperthyroidism and is considered as a diagnostic marker for GD. Therefore, we developed a recombinant protein of human TSHR-169 (hTSHR-169), which was specifically recognized TRAb in the serum of GD patients, and then compare the diagnostic performance between ELISA and dot blot of TRAb tests for their ability to diagnose GD.
20 GD patients and 20 healthy individuals from the Indonesian population were enrolled. TRAb concentration and density were quantified. About Comparative analysis was performed using receiver-operating curve (ROC) analysis.
For dot blot assay, the minimum concentration to detect TRAb requires 100 ng of antigen with antiserum diluted at 1:60.
For diagnosing GD, the ELISA yielded a higher AUC compared with the dot blot assay (0.95 and 0.85, respectively). Using the recommended cutoff values, the efficiency of both assays was examined by comparing the specificity and sensitivity of the assays to the clinical diagnosis.
The ELISA showed 80% and 95%, while the dot blot assay showed 70% and 95% sensitivity and specificity, respectively.
Although the dot blot assay exhibited lower performance than the ELISA method, the dot blot assay is a simple and rapid diagnostic assay that is suitable for diagnosing GD in rural areas, in which healthcare facilities sometimes are not accessible.
| MARK Antibody | |||
| AAT Bioquest | |||
| MARK Antibody | |||
| SAB | |||
| MARK Antibody | |||
| SAB | |||
| MARK Antibody | |||
| EnoGene | |||
| MARK Antibody | |||
| EnoGene | |||
Development and evaluation of indirect antibody ELISA assay for early diagnosis and surveillance of Crimean-Congo hemorrhagic fever infection in humans.
- Crimean-Congo hemorrhagic fever (CCHF) is an important tick-borne zoonotic viral disease of humans.
- CCHF virus causes sporadic cases of severe illness across a huge geographic area across Africa to Europe to Asia including India. CCHF has emerged as a major public health concern in western Indian states including Gujarat and Rajasthan, where regular human cases were reported since the year 2011.
- Humans serve as the dead-end host, and they gain infection via infected tick bite, in close contact with ruminants and from slaughterhouse.
- Currently, the detection of this fatal infection is limited to BSL-4 laboratory which is scarce even in developed economies. Thus, a safe, sensitive assay for early immunodiagnosis is crucial for disease management and containing the outbreak.
- In this study, the conserved recombinant nucleoprotein was exploited as a safe, scalable alternate antigen for the development of indirect IgM and IgG ELISA detection platforms.
- The indirect ELISA was evaluated using suspected clinical samples collected from hotspot areas in India. Comparison with reference MAC ELISA and IgG ELISA revealed a correlation of 95% and 100% respectively. The results indicate that the developed IgM and IgG indirect ELISA has high sensitivity and specificity for detecting CCHFV antibodies among humans.
- These assays are easy to perform and can be employed for high throughput screening of human samples for clinical diagnosis as well as serosurveillance. These assays are also amenable for conversion to low-cost point of care testing formats for application in resource limited settings.
| Recombinant Humanp21 Recombinant Protein | |||
| ProSci | |||
| TWEAK, recombinant / TNFSF12, recombinant (Human) | |||
| PHOENIX PEPTIDE | |||
| TAGLN Recombinant Protein (Rat) (Recombinant- Tag) | |||
| ABM | |||
| TAGLN2 Recombinant Protein (Rat) (Recombinant Tag) | |||
| ABM | |||
| TAGLN3 Recombinant Protein (Rat) (Recombinant Tag) | |||
| ABM | |||
Evaluation of Two Rapid Lateral Flow Tests and Two Surrogate ELISAs for the Detection of SARS-CoV-2 Specific Neutralizing Antibodies.
As vaccination against SARS-CoV-2 progresses rapidly around the world, reliable detection of SARS-CoV-2 specific neutralizing antibodies (NAb) has become an indispensable component of serological diagnostics.
We evaluated the performance of four commercially available tests, i.e. two lateral flow assays (Coris BioConcept COVID-19 Sero NP/RBD and Concile InfectCheck COVID-19 NAb) and two surrogate ELISA (sELISA) tests (EUROIMMUN SARS-CoV-2 NeutraLISA and AdipoGen SARS-CoV-2 Neutralizing Antibodies Detection Kit) in comparison with an in-house SARS-CoV-2 microneutralization test as reference.
| Recombinant Humanp21 Recombinant Protein | |||
| 92-035 | |||
| TWEAK, recombinant / TNFSF12, recombinant (Human) | |||
| 054-85 | |||
| TAGLN Recombinant Protein (Rat) (Recombinant- Tag) | |||
| RP232205 | |||
| TAGLN2 Recombinant Protein (Rat) (Recombinant Tag) | |||
| RP232208 | |||
| TAGLN3 Recombinant Protein (Rat) (Recombinant Tag) | |||
| RP232211 | |||
A total of 334 sera were tested, including 30 samples collected prior to the emergence of SARS-CoV-2, 128 sera from convalescent patients as well as 176 sera from partially or fully vaccinated individuals.
The overall sensitivity of LFAs differed and was 71.6% for the Coris and 98.4% for the Concile. In contrast, overall sensitivity of the NeutraLISA was 86 and 98% for the AdipoGen. All test showed the highest sensitivity when testing samples from fully vaccinated individuals with both sELISA achieving 100% sensitivity. Overall specificity was 89.3% for the Coris and only 58.3% for the Concile.
Similarly, significant differences were observed for both ELISA, with an overall specificity of 82.1% for the NeutraLISA and only 54.8% for the AdipoGen. All tests showed a 100% specificity when testing negative control samples while specificities were lowest when testing samples from only partially vaccinated individuals.
Preparation of an Immunoaffinity Column Based on Bispecific Monoclonal Antibody for Aflatoxin B 1 and Ochratoxin A Detection Combined with ic-ELISA.
- A novel and efficient immunoaffinity column (IAC) based on bispecific monoclonal antibody (BsMAb) recognizing aflatoxin B1 (AFB1) and ochratoxin A (OTA) was prepared and applied in simultaneous extraction of AFB1 and OTA from food samples and detection of AFB1/OTA combined with ic-ELISA (indirect competitive ELISA).
- Two deficient cell lines, hypoxanthine-guanine phosphoribosyl-transferase (HGPRT) deficient anti-AFB1 hybridoma cell line and thymidine kinase (TK) deficient anti-OTA hybridoma cell line, were fused to generate a hybrid-hybridoma producing BsMAb against AFB1 and OTA. The subtype of the BsMAb was IgG1 via mouse antibody isotyping kit test. The purity and molecular weight of BsMAb were confirmed by SDS-PAGE method.
- The cross-reaction rate with AFB2 was 37%, with AFG1 15%, with AFM1 48%, with AFM2 10%, and with OTB 36%. Negligible cross-reaction was observed with other tested compounds. The affinity constant (Ka) was determined by ELISA.
- The Ka (AFB1) and Ka (OTA) was 2.43 × 108 L/mol and 1.57 × 108 L/mol, respectively. Then the anti-AFB1/OTA BsMAb was coupled with CNBr-Sepharose, and an AFB1/OTA IAC was prepared. The coupling time and elution conditions of IAC were optimized.
- The coupling time was 1 h with 90% coupling rate, the eluent was methanol-water (60:40, v:v, pH 2.3) containing 1 mol/L NaCl, and the eluent volume was 4 mL. The column capacities of AFB1 and OTA were 165.0 ng and 171.3 ng, respectively. After seven times of repeated use, the preservation rates of column capacity for AFB1 and OTA were 69.3% and 68.0%, respectively.
- The ic-ELISA for AFB1 and OTA were applied combined with IAC. The IC50 (50% inhibiting concentration) of AFB1 was 0.027 ng/mL, the limit of detection (LOD) was 0.004 ng/mL (0.032 µg/kg), and the linear range was 0.006 ng/mL~0.119 ng/mL.
- The IC50 of OTA was 0.878 ng/mL, the LOD was 0.126 ng/mL (1.008 µg/kg), and the linear range was 0.259 ng/mL~6.178 ng/mL. Under optimum conditions, corn and wheat samples were pretreated with AFB1-OTA IAC. The recovery rates of AFB1 and OTA were 95.4%~105.0% with ic-ELISA, and the correlations between the detection results and LC-MS were above 0.9. The developed IAC combined with ic-ELISA is reliable and could be applied to the detection of AFB1 and OTA in grains.
| anti- Antibody^Polyclonal antibody control antibody | |||
| LSMab09882 | |||
| H2B Antibody Antibody | |||
| E11-184659 | |||
| Lck antibody Antibody | |||
| GWB-250026 | |||
| H2B Antibody Antibody | |||
| MBS8529199-01mg | |||
| H2B Antibody Antibody | |||
| MBS8529199-01mLAF405L | |||
A Commercial Anti-TIF1γ ELISA Is Superior to Line and Dot Blot and Should Be Considered as Part of Routine Myositis-Specific Antibody Testing.
Anti-TIF1γ is an important autoantibody in the diagnosis of cancer-associated dermatomyositis and the most common autoantibody in juvenile onset dermatomyositis.
Its reliable detection is important to instigate further investigations into underlying malignancy in adults. We previously showed that commercial assays using line and dot blots do not reliably detect anti-TIF1γ.
We aimed to test a new commercial ELISA and compare with previously obtained protein immunoprecipitation.
Radio-labelled immunoprecipitation had previously been used to determine the autoantibody status of patients with immune-mediated inflammatory myopathies and several healthy controls. ELISA was undertaken on healthy control and anti-TIF1γ sera and compared to previous immunoprecipitation data.
ZO Rabbit Polyclonal Antibodies |
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| 29274 | SAB | 100ul | 439 EUR |
ABCC2 Rabbit Polyclonal Antibodies |
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| MBS9457710-005mL | MyBiosource | 0.05mL | 300 EUR |
ABCC2 Rabbit Polyclonal Antibodies |
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| MBS9457710-01mL | MyBiosource | 0.1mL | 390 EUR |
ABCC2 Rabbit Polyclonal Antibodies |
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| MBS9457710-5x01mL | MyBiosource | 5x0.1mL | 1610 EUR |
TRPV1 Rabbit Polyclonal Antibodies |
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| MBS9457713-005mL | MyBiosource | 0.05mL | 300 EUR |
TRPV1 Rabbit Polyclonal Antibodies |
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| MBS9457713-01mL | MyBiosource | 0.1mL | 390 EUR |
TRPV1 Rabbit Polyclonal Antibodies |
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| MBS9457713-5x01mL | MyBiosource | 5x0.1mL | 1610 EUR |
TRPA1 Rabbit Polyclonal Antibodies |
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| MBS9457714-005mL | MyBiosource | 0.05mL | 300 EUR |
TRPA1 Rabbit Polyclonal Antibodies |
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| MBS9457714-01mL | MyBiosource | 0.1mL | 390 EUR |
TRPA1 Rabbit Polyclonal Antibodies |
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| MBS9457714-5x01mL | MyBiosource | 5x0.1mL | 1610 EUR |
ABCC2 Rabbit Polyclonal Antibodies |
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| 29220 | SAB | 100ul | 439 EUR |
TRPV1 Rabbit Polyclonal Antibodies |
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| 29223 | SAB | 100ul | 439 EUR |
A total of 110 serum samples were analysed: 42 myositis patients with anti- TIF1γ and 68 autoantibody negative healthy control sera. Anti-TIF1γ was detected by ELISA in 41 out of 42 of the anti-TIF1γ-positive samples by immunoprecipitation, and in none of the healthy controls, giving a sensitivity of 97.6% and specificity of 100%. The false negative rate was 2%.
ELISA is an affordable and time-efficient method which is accurate in detecting anti-TIF1γ.
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2. Gentaur
3. Rockland