Sample
serum, plasma (EDTA, heparin or citrate plasma)
Intended Use
The ELISA test kit provides a quantitative in vitro assay for human antibodies of the immunoglobulin class IgG against tetanus toxoid in serum or plasma (EDTA, heparin or citrate plasma) for the clarification of an unclear immune status.
Contents of Kit
1. Microplate wells coated with antigens, 12 microplate strips each containing 8 individual break-off wells in a frame, ready for use, 12 × 8 STRIPS
2. Calibrator 1, 5 IU/ml (human IgG), ready for use, 1 × 1.5 ml CAL 1
3. Calibrator 2, 2 IU/ml (human IgG), ready for use, 1 × 1.5 ml CAL 2
4. Calibrator 3, 1 IU/ml (human IgG), ready for use, 1 × 1.5 ml CAL 3
5. Calibrator 4, 0.1 IU/ml (human IgG), ready for use, 1 × 1.5 ml CAL 4
6. Calibrator 5, 0.01 IU/ml (human IgG), ready for use, 1 × 1.5 ml CAL 5
7. Positive control (IgG, human), ready for use, 1 × 1.5 ml
8. Negative control (IgG, human), ready for use, 1 × 1.5 ml
9. Enzyme conjugate, peroxidase-labelled anti-human IgG, ready for use, 1 × 12 ml
10. Sample buffer, ready for use, 1 × 100 ml
11. Wash buffer, 20x concentrate, 1 × 50 ml
12. Substrate solution, TMB/H2O2, ready for use, 1 × 12 ml
13. Stop solution, 0.5 M sulfuric acid, ready for use, 1 × 12 ml
14. Protective foil --- 2 pieces of FOIL
15. Test instructions --- 1 booklet
16. Quality control certificate --- 1 protocol
Storage
The test kit has to be stored at a temperature between +2°C to +8°C. Do not freeze. Unopened, all test kit components are stable until the indicated expiry date.
Performance Characteristics
Calibration: The Anti-Tetanus Toxoid ELISA (IgG) was calibrated using the 2nd international WHO standard preparation 13/240 (WHO International Standard 2nd International Standard for Anti-Tetanus Immunoglobulin Human NIBSC code: 13/240).
For every group of tests performed, the extinction values of the calibrators and the international units determined for the positive and negative controls must lie within the limits stated for the relevant test kit lot. A quality control certificate containing these reference values is included. If the values specified for the controls are not achieved, the test results may be inaccurate and the test should be repeated. The binding activity of the antibodies and the activity of the enzyme used are temperature-dependent. It is therefore recommended using a thermostat in all three incubation steps. The higher the room temperature (+18°C to +25°C) during the incubation steps, the greater will be the extinction values. Corresponding Variations also apply to the incubation times. However, the calibrators are subject to the same influences, with the result that such variations will be largely compensated in the calculation of the result.
Antigen: The microtiter wells were coated with inactivated tetanus toxoid antigen.
Precision
Intra-assay variation ＜5%
Inter-assay variation ＜10%
Detection Limit
The lower detection limit is defined as the mean value of an analyte-free sample plus three times the standard deviation and is the smallest detectable antibody titer. The lower detection limit of the Anti-Tetanus Toxoid ELISA (IgG) is 0.001 IU/ml.
General Description
The Anti-Tetanus Toxoid ELISA (IgG) is based on inactivated tetanus toxin and is designed for the
quantitative determination of human IgG antibodies against tetanus toxoid in serum or plasma. This test is suited for both investigation the immune status and for vaccination control.
Tetanus vaccine, also known as tetanus toxoid (TT), is a toxoid vaccine used to prevent tetanus. During childhood, five doses are recommended, with a sixth given during adolescence.
After three doses, almost everyone is initially immune, but additional doses every ten years are recommended to maintain immunity. A booster shot should be given within 48 hours of an injury to people whose immunization is out of date.
Confirming that pregnant women are up to date on tetanus immunization during each pregnancy can prevent both maternal and neonatal tetanus. The vaccine is very safe, including during pregnancy and in those with HIV/AIDS.
Redness and pain at the site of injection occur in between 25% and 85% of people. Fever, feeling tired, and minor muscle pain occurs in less than 10% of people. Severe allergic reactions occur in fewer than one in 100,000 people.
A number of vaccine combinations include the tetanus vaccine, such as DTaP and Tdap, which contain diphtheria, tetanus, and pertussis vaccines, and DT and Td, which contain diphtheria and tetanus vaccines. DTaP and DT are given to children less than seven years old, while Tdap and Td are given to those seven years old and older. The lowercase d and p denote lower strengths of diphtheria and pertussis vaccines.
Tetanus antiserum was developed in 1890, with its protective effects lasting a few weeks. The tetanus toxoid vaccine was developed in 1924, and came into common use for soldiers in World War II. Its use resulted in a 95% decrease in the rate of tetanus. It is on the World Health Organization's List of Essential Medicines.
Standard Curve
The following plot is an example of a typical calibration curve. Please do not use this curve for the determination of antibody concentrations in patient samples.
Citations
Publication ()
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Bioengineered human tonsil organoids as an immuno-engineering platform for evaluating immune functions
Intan Rosalina Suhito, Donovan Eu Kum Chuen, Hong Sheng Cheng, Nguan Soon Tan, Zhe Zhang Ryan Lew, Kai Sen Tan, Justin Jang Hann Chu, Giselle G.K. Ng, Yvonne C.F. Su, Gavin J.D. Smith i, Chee Wah Tan, and Andy Tay
Applications: ELISA
Reactive species: Tetanus Toxoid
"Abstract: Although animal models remain the preclinical gold standard, they are constrained by interspecies differences in adaptive immunity, including antibody class-switching, affinity maturation, and
multicellular interactions. To address this gap, we established a more physiologically relevant platform, a bioengineered tonsil organoid (BTO) system that combines autologous tonsil-derived
stromal cells with immune cells to generate highly uniform organoids, preserving multicellular complexity and sustained stromal-immune interactions. BTO cultures mounted robust, antigenspecific recall responses to influenza, tetanus, and COVID-19 antigens. Notably, stimulation with ovalbumin and SARS-CoV-1 spike protein expanded antigen-recognizing B cells, indicating
capacity for naïve responses. Digital spatial transcriptomics and proteomics analysis confirmed immune-stromal crosstalk underpinning both humoral and cellular responses of BTO after
challenge. These data highlight stromal cells as active scaffolds that not only maintain 3D architecture but also facilitate immune activation, memory recall, and recognition of naïve antigens. Together, BTO offers a human-relevant model for mechanistic immunology and for pre-clinical evaluation on immunotherapies and vaccines not achievable with 2D cultures or animal models."
Article snippet: For tetanus toxoid-specific IgG detection, culture supernatants were assayed by human tetanus toxoid IgG ELISA kit (Creative Diagnostics) following the manufacturer’s instructions. 
Figure 1. Tonsil organoids mount effective memory recall responses to previously exposed antigens.
COA
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