Immuno-Diagnostics Antibodies & Antigens for Veterinary NCDs (noncommunicable diseases) Test

GeneMedi’s anti-SAA monoclonal antibodies demonstrate superior performance in immunoassay.

GeneMedi’s monoclonal antibodies for SAA (Serum Amyloid A) offer high sensitivity for detecting SAA in cat, dog, and horse serum. Due to the high sequence homology among these species, the same antibody pairs can be used to develop versatile sandwich immunoassays. As a result, GeneMedi’s anti-SAA antibodies provide a valuable tool for accurately monitoring and diagnosing inflammatory conditions across multiple species.

Figure 1. GeneMedi’s monoclonal antibodies demonstrate high sensitivity and specificity for detecting SAA in both healthy and inflammatory cat plasma samples using a sandwich immunoassay.

A. Calibration curve for GMP-FEL-Serum-amyloid-A-Ag01 in mixed cat plasma in the sandwich immunoassay. The SAA concentration in mixed cat plasma was 145 μg/ml, and the sample was diluted 208 times prior to testing.
B-C. Dilution curves for recombinant cat SAA antigen and plasma samples using the prototype reagents in the sandwich immunoassay. (B) Healthy cat samples (H1, H2), with SAA concentrations of 0.67 μg/ml and 0.45 μg/ml, respectively. Plasma samples were diluted 45 times before testing. The initial concentration of GMP-FEL-Serum-amyloid-A-Ag01 was 65 ng/ml. (C) Postoperative cat samples (P1, P2), with SAA concentrations of 169 μg/ml and 220 μg/ml, respectively. Plasma samples were diluted 200 times before testing. The initial concentration of GMP-FEL-Serum-amyloid-A-Ag01 was 200 ng/ml.
D. SAA concentrations in healthy and postoperative inflammatory cats. Healthy cat samples were pre-diluted 100-3000 times, while inflammatory cat samples were pre-diluted 9000 times prior to testing.

Figure 2. GeneMedi’s anti-SAA monoclonal antibodies enable sensitive detection of SAA in dog, horse, and cat serum using a sandwich immunoassay.

A-B. Dilution curves of recombinant dog (A) and horse (B) SAA antigens in the sandwich immunoassay. GMP-CAN-Serum-amyloid-A-Ab01 was coated and blocked at 37°C for 1 hour, and incubated with GMP-CAN-Serum-amyloid-A-Ag01 and GMP-EQU-Serum-amyloid-A-Ag01 at 37°C for 1 hour.
C. The recommended antibody pairs.
D-E. Comparison of SAA detection results between diseased and healthy individuals in dogs (D) and horses (E) in the sandwich immunoassay. Healthy serum was pre-diluted 50 times, while diseased serum was pre-diluted 1500 times (horse) or 2500 times (dog), with the dilution buffer containing 1% casein and 0.05% Tween 20.
F-H. Testing results of SAA in cat (F), dog (G), and horse (H) serum samples in the sandwich immunoassay. GMP-FEL-Serum-amyloid-A-Ab10 was blocked for 1 hour (blocking solution containing 2.5% sodium caseinate). Dog serum was pre-diluted 2500 times, while cat and horse serum were pre-diluted 1500 times, using a Tris buffer containing 0.01% CHAPS.

Immunoassay calibration curve for NT-proBNP

These immunoassays have a sensitivity of 22 pg/mL for measuring recombinant NT-proBNP. Figures 1 and 2 display the suggested pairing calibration curves. Conversion of 1NT-proBNP concentration: 1 pmol/L = 1÷10.545 pg/ml Please note that a variety of factors might affect how well an immunoassay test performs. These consist of the labeling procedure, the kind of label coupled to the detection antibody, and the detection platform. Thus, on our customers' immunoassay systems, other NT-proBNP antibody pairs with non-overlapping epitopes could perform better.

Figure 1-2. NT-proBNP immunoassay calibration curve

(left) Calibration curve for optimal paired detection
(right) Calibration curve and canine plasma sample dilution comparison

Reagent type: two-step sandwich immunofluorescence detection, streptavidin-coated microplate
Capture antibody: 200ng/well, conjugated biotin
Capture antibody: 200ng/well, labeled europium chelate
Antigen: Canine recombinant NT-proBNP (Cat.#GMP-CAN-NT-proBNP-Ag01)
Sample volume: 50μl
Incubation time: 40 minutes at room temperature

Sensitivity detection of NT-proBNP

These immunoassays have a sensitivity of 22 pg/ml 1 for measuring recombinant NT-proBNP. Figures 1 and 2 display the suggested pairing calibration curves. Conversion of 1 NT-proBNP concentration: 1 pmol/L = 1÷10.545 pg/ml Please be aware that a variety of factors might affect how well an immunoassay test performs. These consist of the labeling procedure, the kind of label coupled to the detection antibody, and the detection platform. Thus, on our customers' immunoassay systems, other NT-proBNP antibody pairs with non-overlapping epitopes could perform better.

Figure 3. NT-proBNP concentration in EDTA plasma samples from healthy dogs and dogs with heart disease.

Reagent type: two-step sandwich immunofluorescence detection, streptavidin-coated microplate
Capture antibody: GMP-CAN-NT-proBNP-Ab01, 1μg/well
Capture antibody: GMP-CAN-NT-proBNP-Ab02, 200ng/well, labeled europium chelate
Calibrator: Canine recombinant NT-proBNP (Cat.# GMP-CAN-NT-proBNP-Ag01)
Sample volume: 50μl
Incubation time: 40 minutes at room temperature

Stability Test of NT-proBNP

Our antibodies were tested for their ability to detect endogenous canine NT-proBNP during sample storage. EDTA plasma from dogs with heart disease was incubated at two temperatures: +4°C and +20°C. At +4°C, NT-proBNP remains stable for at least 72 hours (95%–105% of initial immunoreactivity, Figure 4A). Simultaneously, at +20°C, about 98% of the initial immunoreactivity is detectable after 24 hours (Figure 4B). Using our recommended antibody pairings, plasma can be stored at +4°C for at least 72 hours with minimal loss of immunoreactivity. However, at room temperature, there is a slight weakening of the signal during the first 24 hours. Note that individual plasma or serum samples may vary.

Figure 4-5. Stability of endogenous canine NT-proBNP in EDTA plasma

(left) Immunoreactivity of NT-proBNP in plasma samples incubated for 24, 48 and 72 hours at +4°C.
(right) Immunoreactivity of NT-proBNP in plasma samples incubated for 4, 8, 24 and 48 hours at +20°C.

Collect protease inhibitor-free EDTA plasma; samples are centrifuged, separated, and stored at -70°C before use. EDTA plasma is stored at +4°C or +20°C with the addition of 0.1% NaN3 to prevent bacterial growth. After incubation, samples were stored at -70°C before measurement. The NT-proBNP concentration in plasma was 9 ng/ml (determined by GMP-CAN-NT-proBNP-Ab01 and GMP-CAN-NT-proBNP-Ab02 immunoassays). Detection using GMP-CAN-NT-proBNP-Ab01 and GMP-CAN-NT-proBNP-Ab05 as capture antibodies and GMP-CAN-NT-proBNP-Ab06 and GMP-CAN-NT-proBNP-Ab02 as detection antibodies respectively The system results are depicted in Figure 3.

GeneMedi's feline cTnI antibodies exhibit superior performance in sandwich immunoassays.

GeneMedi’s GMP-FEL-cTnI-Ab01, GMP-FEL-cTnI-Ab02, and GMP-FEL-cTnI-Ab05 demonstrate superior performance in sandwich ELISA assays. We tested four antibody pairs, and the GMP-FEL-cTnI-Ab01 and GMP-FEL-cTnI-Ab02 pair showed the lowest EC50. Based on these results, we recommend this antibody pair for optimal performance in cTnI detection.

Figure 1. The performance of GeneMedi’s feline cTnI antibodies in sandwich ELISA assays.

A-B. Results of four different antibody pairs in sandwich ELISA assays.
C. Antibody pairs and their EC50 values.