Meridian is a primary manufacturer of specialized high-quality molecular reagents and offers solutions to a wide range of industries to diagnose and treat diseases, discover new therapeutics or develop tests for environmental, food and cosmetic safety.
New HCV Chimeric Antigen (NS3/Core/NS4/NS5) Reacts with HCV-positive serum in lateral flow assays
Globally, an estimated 58 million people have chronic hepatitis C virus (HCV) infection, with about 1.5 million new infections occurring per year. Direct-acting antiviral medicines (DAAs) can cure more than 95% of HCV infections, but access to diagnosis and treatment is low. Because HCV infections are usually asymptomatic, they are not detected until decades have passed and symptoms develop due to serious liver damage caused by the virus. HCV diagnosis commonly involves two stages: (1) Testing for anti-HCV antibodies with a serological test and (2) confirmation by molecular testing . Most commercial anti-HCV tests that can detect seroconversion at 8 weeks include the core antigen and antigens to NS3, NS4 and NS5 proteins. However, these tests require a blend of individual antigens – these antigens can have varying affinity to HCV antibodies which can cause assay reproducibility issues. Meridian’s new HCV chimeric contains four antigens expressed in E. coli (NS3, Core, NS4, NS5) and demonstrates high reactivity with HCV-positive serum in lateral flow assays.
E1/E2 Glycoproteins
Capsid
RNA
Core
Envelope
Hepatitis C virus
• E. coli expressed chimeric containing HCV core, NS3, NS4 and NS5 • M olecular weight of 33 kDa as measured by SDS-PAGE • C ontains N-term His-Tag • Specifically designed to work as a pair in a lateral flow or ELISA antibody detection assays HCV Chimeric NS3/Core/NS4/NS5 (Subtype 1b), Cat #BN1250
6His
NS3
Core
NS4
NS5
HCV Chimeric Antigen
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Performance Data
Fig 1 . SDS-PAGE analysis of HCV chimeric antigen SDS-PAGE was performed with a 10% Acrylamide gel, followed by staining with an InstantBlue Coomassie protein stain. Purified HCV chimeric antigen (Cat# BN1250) showed a single band with a molecular weight of 33kDa. The purity of HCV chimeric antigen was observed to be approximately >90% using ChemiDoc densitometric scanning.
2a
2b
MLS Cat# BN1250
100% R21
88% R20
80% R19
70% R18
60% R17
54% R16
50% R15
HCV Recombinant Antigen - Gold conjugated
46% R14
40% R13
34% R12
30% R11
Anti-HCV Ab (Patient’s serum)
26% R10
20% R9
16% R8
14% R7
12% R6
Anti-Human IgG
10% R5
7% R4
Membrane
5% R3
3% R2
1% R1
Fig 2. Rapid Lateral Flow Immunochromatography Test using HCV chimeric antigen BN1250 HCV Antibody Test assay was developed with two pre-coated lines, “C” Control line, and “T” Test line on the surface of the nitrocellulose membrane. Anti-Chicken IgY was coated on the control line, and anti Human IgG was coated on the test line region as per the figure (2a). The recombinant HCV chimeric antigen Cat# BN1250 was conjugated with colloidal gold particles and used as a detector for HCV antibodies. During the test, HCV antibodies in the patient’s serum sample interacted with recombinant HCV antigen conjugated with colloidal gold particles to form antibody-antigen gold particle complex. This complex migrated on the membrane via capillary action until the test line, where it was be captured by the monoclonal anti-human IgG. (2b). The visual limit of detection (LoD) as defined by the lowest dilution range on T line color to see be visible to the naked eye, was 2-9 for Cat# BN1250.
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Email: info@meridianlifescience.com Orders: orders@meridianlifescience.com www.MeridianBioscience.com/lifescience
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11/23
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