Relative Reduction in RF Interference in CA19-9 Assay
Performance Comparison – TRU Block vs Competitors (Patient Sample: HAMA Serum 61)
0 0.2 0.4 0.6 0.8 1 1.2
3,5
3,0
2,5
2,0
1,5
No Blocker
TRUBlock
Competitor
Mouse IgG
Rabbit IgG
1,0
No Blocker
5µg/mL
2.5µg/mL
1.25µg/mL
0,5
Figure 2: A double mouse monoclonal sandwich CA19-9 assay was used to measure the effectiveness of TRU Block, Mouse IgG and a competitor HA blocker in blocking RF interference from a commercially sourced patient sample (A12916H). RF activity was measured in the absence (no blocker) and in the presence of blockers (TRU Block, Competitor, Mouse IgG and Rabbit IgG) at various concentrations (1.25 mg/ml, 2.5 mg/ml and 5 mg/ml). Greater suppression of the RF signal with no blocker ( blue ) indicates greater RF blocking effectiveness.
0,0
Competitor A
Competitor B
TRU Block
(Patient Sample: HAMA Serum 69)
3,5
3,0
2,5
hindrance mechanism. Performance advantages of TRU Block include broader coverage against all types of HA inference and RF, the ability to be used at a low concentration, and better blocking efficacy compared to Mouse IgG. The blocking effectiveness of TRU Block has been evaluated against Mouse IgG and a well-known HA blocker using both two-step (Figure 1) and one-step (Figure 2) double mouse monoclonal ELISAs with blockers added to the sample diluent buffer. The results indicate that TRU Block can outperform both Mouse IgG and the other blocker in ELISA-based immunoassays. TRU Block has also been successfully used in chemiluminescent assays and lateral flow rapid tests where TRU Block was dried down as a stripe between the sample pad and assay antibody location.
SUMMARY Heterophilic antibody interference blockers are an essential part of clinical diagnostic assays, and their importance has been documented by dozens of case studies where approved assays resulted in misdiagnosis (Bolstad, N. et al, 2013). In selecting a blocker, it is important to consider the source of the antibodies used in the assay and the types of heterophilic interference that could impact the assay. TRU Block™ is a unique HA, HAMA and RF interference blocker that can be used in ELISA, LF and chemiluminescent assay formats. It has several performance advantages over other heterophilic antibody interference blockers on the market and when used in an immunoassay, can prevent false positive and false negative results.
2,0
1,5
1,0
0,5
0,0
Competitor A
Competitor B
TRU Block
(Patient Sample: HAMA Serum 70)
3,5
3,0
2,5
2,0
1,5
1,0
0,5
0,0
Competitor A
Competitor B
TRU Block
Bolstad, N., Warren, D. J. & Nustad, K. Heterophilic antibody interference in immunometric assays. Best Pract Res Clin Endocrinol Metab 27, 647–661 (2013).
250µg/mL
125µg/mL
62.5µg/mL
31.25µg/mL
No blocker
Figure 3: Customer (IVD manufacturer) One-step ELISA results: HAMA interference for three different samples (HAMA Serum 61, Hama Serum 69 and HAMA Serum 70) was measured initially in the absence of blockers to determine 100% interference signal level. HAMA activity was then measured in the presence of blockers (added with assay antibodies together) to measure the suppression of signal. Greater suppression of signal (reduced bar height) indicates a more effective HAMA blocker.
Product
Cat Number Protein Concentration
Application ELISA & LF
TRU Block Ready 8001
Single-step dilution with recommended dilution of 1:1000 to 1:10
TRU Block ULTRA 8000
Range: 24 - 26 mg/mL
ELISA, CLIA & LF
TRU Block TRU Block 2 TRU Block 3
A66800H Range: 24 - 26 mg/mL A66802H Range: 24 - 26 mg/mL
ELISA
ELISA, CLIA & LF
A66803H 24.3 mg/mL
ELISA, CLIA
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