Sensitive CG-ICA based on heterologous coating antigen and mAb prepared with carbons-linker immunogen

Figures & data

Figure 1. The OD value and inhibition ratio of ic-ELISA with three coating antigens and three antibodies from three immunogens. (A) The synthesis of haptens, hapten-conjugations and the process of immunizing mice. (B) The OD value and inhibition ratio were evaluated using the ic-ELISA.

Table 1. Competitive ELISA results with different coating antigens and antibodies

Immunogen	Coating antigen	Coating antigen (μg/mL)	Dilution of antiserum	Titer (OD450)	Inhibition (%)
H1-BSA	H1-OVA H2-OVA H3-OVA	0.3 0.3 0.3	1/3000 1/3000 1/3000	0.572 0.714 0.812	10.57 12.67 10.35
H2-BSA	H1-OVA H2-OVA H3-OVA	0.3 0.3 0.3	1/3000 1/3000 1/3000	0.137 1.453 1.197	5.18 6.90 17.92
H3-BSA	H1-OVA H2-OVA H3-OVA	0.3 0.3 0.3	1/3000 1/3000 1/3000	0.527 1.628 1.677	16.23 63.60 48.54

Figure 2. The standard curves of the ic-ELISA for detection of AMD with the five monoclonal antibodies. Each point of inhibition curve represents three replicates.

Figure 3. Specificity analysis of the ic-ELISA. The concentrations of amantadine, rimantadine, 1-adamantane carboxylic acid, ribavirin, paracetamol, danofloxacin, and olaquindox in PBS were 100 ng/mL.

Figure 4. Standard curve of CG-ICA for detection of AMD obtained by plotting the logarithm of B/B₀ against the various AMD concentrations. Error bars represent the standard deviation (n = 3).

Table 2. Recovery and CV of CG-ICA in chicken samples.

Spiked level (μg/kg)	Mean (μg/kg)	Recovery (%)	Intra-day CV (%)^a	Inter-day CV (%)^b
0.5	0.615	123.0	3.5	6.7
1.0	1.203	120.3	2.4	5.9
2.5	2.410	96.4	5.8	9.2

^a The intra-day CV was completed in one day with seven replicates at each concentration.

^b The inter-day CV was completed every day for four consecutive days with three replicates at each concentration.

Data availability

The data used to support the findings of this study are included in the article.