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.
Sustainable Solutions for Immunoassays Animal-free Blockers for a Resilient Tomorrow Sustainable novel reagent solutions for diagnostic testing are at the forefront of modern healthcare innovation. These reagents are designed with environmental responsibility in mind, minimizing waste generation during their production. They are eco-friendly alternatives that reduce the ecological footprint of diagnostic assays, utilizing animal-free processes and green chemistry principles while making healthcare more environmentally sustainable. Meridian is committed to supporting diagnostic innovation with sustainable solutions that also enhance patient care.
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Animal-Free Blockers
Immunoassay interference blockers are reagents used in Enzyme Linked Immuno Sorbent Assays (ELISA) and Lateral Flow (LF) assays to reduce interference from proteins in patient samples that could produce false results and an incorrect diagnosis. Non-specific interactions occur between antibodies within the assay itself and endogenous antibodies from the patient’s sample such as heterophilic antibodies and autoantibodies. HAMA (Human anti-mouse antibodies), heterophilic antibodies (HA) and rheumatic factor (RF) and can cause significant interferences in immunoassays leading to false-positive or false-negative results.
Immunoassay interference blockers are usually produced using animal sources which can lead to batch-to-batch variation, low specificity and interference from cross-reactive proteins — all of which can compromise assay performance. Recombinant technologies have enabled animal-free alternatives that offer greater reproducibility, scale-up and overall alleviate ethical concerns. Since 2019, the European Union Directive 2010/63/EU encourages the use of animal-free reagents. Companies that truly care about sustainability and the environment are encouraging scientists to minimize the use of animal-derived products in their work.
Meridian’s new blockers, Mouse-Free IgG and K-Block ™ are 100% animal-free.
WHY GO ANIMAL FREE?
Enhanced specificity: Due to their higher purity and homogeneity, animal-free blockers offer improved specificity over traditional blockers, as they will not cross-react with animal-sourced antigens and antibodies in the assay. The potential for non-specific binding is reduced and the signal-to-background ratio is improved. Consistent performance: Animal-derived reagents can have batch-to-batch variability, impacting assay consistency. In contrast, animal-free reagents are more chemically well-defined, resulting in regulatory advantages and quicker time-to-market.
Ethical considerations: Using an animal protein-free blocker aligns with the global ethical concerns on animal welfare and the reduction of animal usage in science. Regulatory compliance: Regulations such as the European Union (EU) Directive 2010/63/EU, are encouraging the use of animal-free reagents. Reduced risk of contamination: Animal-derived components used in traditional blockers can contain carry-over contaminants, such as viruses, prions, and endotoxins, directly affecting the accuracy of an assay. Animal-free blocker eliminates this risk.
Cruelty free Animal free
Same HAMA blocking potency as native version
Sustainable commercial-scale manufacturing
Economically Feasible
Supply continuity
Mouse-FREE IgG (Recombinant, Animal-Free HAMA & HA Blocker)
Meridian Bioscience’s Mouse-FREE IgG is a HAMA blocker made from a mixture of full-length recombinant antibodies identified in native mouse IgG (IgG1, IgG2a, IgG2b, IgG3). Mouse-FREE IgG is a direct replacement to the animal version (native mouse IgG) and it has the same HAMA blocking efficiency (Fig 1).
Mouse-FREE IgG Cat# BN1300
HAMA Blocking Effectiveness - Meridian Mouse-FREE IgG vs. Native Mouse Serum
Blocking antibodies such as mouse IgG neutralize the interfering antibodies present in serum samples resulting in accurate binding of the analyte of interest by the capture and detection antibodies. Human HAMA-positive serum was incubated with Meridian’s Mouse-FREE IgG (Cat No. BN1300) or serum-derived purified native mouse IgG at concentrations ranging from 0.0001-1.25 μ g/mL for 20 mins at room temperature. The effectiveness of each blocker was determined by comparing the relative suppression of the HAMA signal – the lower EC50 values indicate greater interference suppression.
Mouse IgG
0 0.005 0.01 0.015 0.02 0.025 0.03 0.035
Mouse IgG
Interfering HA (e.g. HAMA)
IgM RF
Detection Antibody (mouse origin)
Antigen
Blocking Buffer
Capture Antibody (mouse origin)
Meridian Mouse-FREE IgG
Native Mouse IgG
A) HAMA Interference
100
100
100
80
80
69
66
60
60
42
40
40
20
20
0
0
Native Mouse IgG
Competitor Blocker #1
Competitor Blocker #2
Meridian Mouse-Free IgG
0.0001
0.001
0.01
0.1
1
10
Blocker concentration ( µ g/ml)
B) RF Interference
100
100
100
80
80
60
60
40
40
20
20
14
14
11
0
0
0.01
0.1
1
10
100
Meridian Mouse-Free IgG
Native Mouse IgG
Competitor Blocker #1
Competitor Blocker #2
Blocker concentration ( µ g/ml)
Meridian Mouse-Free IgG | Native mouse IgG | Competitor Blocker #1 | Competitor Blocker #2
Fig 1. A double mouse monoclonal sandwich assay was used to compare the performance of Meridian’s Mouse-FREE IgG (100% animal-free, recombinant Mouse IgG, Cat No. BN1300) against serum-derived purified native mouse IgG and two competitor HAMA blockers. (A) Human HAMA-positive or (B) RF-positive serum was preincubated with the individual blockers for 20 minutes at room temperature. Blocker concentrations ranged from 0.0003-1.25 μ g/mL for the HAMA assay and 0.0195-80 μ g/mL for the RF assay. The effectiveness of each blocker was determined by comparing the relative suppression of the HAMA or RF signal – lower EC50 values indicate greater interference suppression. The results demonstrate that Mouse-Free IgG is significantly more effective than native mouse IgG and the two competitor blockers at blocking HAMA and RF interference.
www.meridianbioscience.com/lifescience
K-Block TM Recombinant, 100% Animal-free HAMA, RF and HA All-in-One Blocker
Meridian Bioscience’s newest solution, K-Block TM is a HAMA/RF/HA interference blocker that is 100% animal-free and designed for ELISA, chemiluminescent and lateral flow immunoassays. It is the first commercially available, animal-free heterophilic interference blocker that has proven high performance against similar animal-based blockers (Fig 2).
K-Block TM Cat# BN1200
PERFORMANCE DATA K-Block TM demonstrates superior HAMA and RF blocking over Mouse IgG and other commercial HAMA blockers.
A.
B.
Relative HAMA Blocking Capacity
Relative RF Blocking Capacity
100
100
Meridian K-Block TM Native mouse IgG Competitor Blocker #1 Competitor Blocker #2
Meridian K-Block TM Native mouse IgG Competitor Blocker #1 Competitor Blocker #2
80
80
60
60
40
40
20
20
0
0
0.0001
0.001
0.01 Blocker Concentration ( µ g/ml)
0.1
1
0.01
0.1
Blocker Concentration ( µ g/ml) 1
10
100
HAMA Blocking Strength Relative to K-Block TM
RF Blocking Strength Relative to K-Block TM
100
100
100.0
100.0
80
80
60
60
50.0
45.6
40
40
20
20
22.8
10.3
9.9
7.7
0
0
Meridian K Block TM
Native Mouse IgG
Competitor Blocker #1
Competitor Blocker #2
Meridian K Block TM
Native Mouse IgG
Competitor Blocker #1
Competitor Blocker #2
Fig 2. A double mouse monoclonal sandwich assay was used to compare the performance of Meridian Bioscience’s K-Block ™ (100% animal free, recombinant blocker, MLS Cat No. BN1200) against serum-derived purified native mouse IgG and two competitor’s recombinant mouse IgG HAMA blockers. (A) Human HAMA-positive or (B) RF-positive serum was preincubated with the individual blockers for 20 minutes at room temperature. Blocker concentrations ranged from 0.0001-1.25 μ g/mL for the HAMA assay and 0.01-80 μ g/mL for the RF assay. The effectiveness of each blocker was determined by comparing the relative suppression of the HAMA signal to the signal of samples with no blocker. The results demonstrate that at higher concentrations K-Block is as effective as the other blockers and at lower concentrations, K-Block was more effective than native mouse IgG and the two competitor’s recombinant HAMA blockers.
Ordering information: USA
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