Biomat Developed Coated Plates for Immunotoxicity ELISA Tests

Immunotoxicity Plates for ELISA Tests

Recent studies indicate that the immune system of humans and animals is vulnerable to adverse effects from drugs, environmental chemicals, and biotechnology products, potentially leading to morbidity and mortality.

Immunotoxicology is the study of the harmful effects of xenobiotics on the immune system. Adverse responses can include:

Increased susceptibility to infections and tumorigenesis
Hypersensitivity reactions
Autoimmune diseases

Regulatory agencies such as the EPA, FDA, EMEA, and Japan’s MHLW emphasize evaluating potential immunotoxicity in the immune system.

One key test, the T-cell-dependent antibody response (TDAR), is recognized for its predictive value in detecting drug-induced immunotoxicity. Using exogenous antigens, this test assesses overall immune function and forecasts potential immunoregulatory effects.

Biomat’s Coated Plates for Immunotoxicity ELISA Tests

ELISA (enzyme-linked immunosorbent assay) is a powerful method to measure antibodies, allowing serial sample collection and reducing reagent usage. Biomat offers 96 Well Plates for Immunotoxicity detection with specific antigens:

KLH (Keyhole Limpet Hemocyanin): used as vaccine carrier protein acting as the hapten carrier part of the vaccine component or as a highly immunogenic antigen to assess the immune competence of an organism and as a carrier of low molecular mass peptide and haptens, such as oligosaccharides, gangliosides or (glyco)peptides, designed to facilitate antibody production- KLH Coated Plates
Tetanus Toxoid: commonly called lockjaw, is a serious bacterial disease that affects muscles and nerves. It is characterized by muscle stiffness that usually involves the jaw and neck that then progresses to involve other parts of the body. This disease is caused by neurotoxin from deep wound infection with Clostridium tetani. Tetanus toxoid Coated Plates can be used for assays where the level of anti-tetanus toxoid antibodies present in biological samples are measured spectrophotometrically.
DNP (Dinitrophenol) and TNP (Trinitrophenol): are haptens that can be attached to carrier proteins such as KLH (Keyhole Limpet Hemocyanin) or ovoalbumin. When one of these complexes is injected into appropriate animal models, produce an immune response which is measured as changes in the levels of anti DNP (or anti TNP) IgM and IgG. Thanks to these changes, the researchers can assess the impact of pharmacologic or genetic manipulations on the studied immune system. DNP Coated Plates and TNP Coated Plates

Why Choose Biomat?

Biomat’s 30+ years of expertise in coating development ensure high-quality, reliable products. Our coated plates are ideal for ELISA IgM and IgG assays, commonly used as biomarkers of immunotoxicity.

Uses in Preclinical Studies

These Coated Plates for Immunotoxicity ELISA test are ideal for the setup of ELISA IgM and IgG assays to be used as biomarkers of immunotoxicity because they measure levels of IgM and IgG of anti KLH, anti DNP, anti TNP and anti-Tetanus toxoid in your samples.

Anti-KLH, anti-tetanus toxoid, anti-DNP and anti-TNP IgM and IgG are routinely used as biomarkers of immunotoxicity.

In preclinical studies, animals are immunized with an antigen such as KLH while being dosed with a drug candidate. The levels of anti-KLH IgM and IgG are determined in serum or plasma collected at 5-7 or 14-21 days and compared with those in a control group that was not exposed to the drug. A decrease in anti-KLH levels in the treatment group provides evidence of immunosuppression.

Example Protocol for Mouse Anti-IgG KLH Determination

Add 100 µl of different concentrations of monoclonal mouse IgG anti KLH (BioLegend code 408502 at 0.5 mg/ml, from 0.0005 to 0.500 µg/ml, diluted in Sample Diluent) to the wells of KLH coated plate and incubate for 60 minutes at room temperature
Empty the wells and wash with Wash Buffer four times
Add 100 µl /well of goat anti-mouse IgG -HRP (Jackson ImmunoResearch code 115-035-003, diluted 1:25,000 in Diluent for HRP conjugate) and incubate for 60 minutes at room temperature
Empty the wells and wash with Wash Buffer four times
Add 100 µl /well of TMB substrate solution) and incubate 15 minutes at room temperature
Stop the substrate reaction by adding 100 µl /well of Stop Solution for substrate and read the optical density values at 450 nm

Figure 1: the figure shows the mO.D. directly proportional to the concentration of mouse IgG anti KLH bound to KLH coated microplate.

Figure 2: the figure gives an idea of the dilution factor to apply to the serum/plasma of the immunized mouse under evaluation; where k means a dilution of 1:1,000.