Mycotoxins are toxic metabolites produced by molds, which have strong carcinogenicity and reduce immunity. Common mycotoxins mainly include aflatoxin, ochratoxin, DON, trichothecenes (T-2 toxin), zearalenone, fusarium moniliformin, fumonisins, etc. Grain and feed are easily contaminated by mycotoxins, and this problem has increasingly become a major problem that cannot be ignored in modern agricultural production.
Welch Materials has developed a series of mycotoxin immunoaffinity column products with its strong scientific research strength and advanced production technology. Applicable to various testing laboratories, import and export institutions, etc.
Generally, a complete immunoaffinity column consists of 6 parts:
• Column tube (1mL/3mL)
• Stationary phase (agarose gel)
• Antibodies (agar-coupled specific antibodies)
• Protective solution (usually a buffer solution to keep the agar wet)
• frit and top and bottom caps (to prevent column fluid loss)
Immunoaffinity column technology is immunoaffinity chromatography technology, which is a purification technology that simplifies the sample extraction process and effectively improves the purity of the sample in the process of sample pretreatment. Due to its strong specificity, it belongs to trace detection in food analysis, and the detection limit very low. Welchom® immunoaffinity columns are used to separate and purify mycotoxins from samples based on the specific reaction between antigens and antibodies. Antibodies in the immunoaffinity column are suspended in the gel by covalent bonding and specifically adsorb the mycotoxins in the sample. If the sample for detection contains mycotoxins, the toxins are captured and bound by the antibody as the sample passes through the immunoaffinity column. All other substances are washed from the immunoaffinity column. Methanol was used as the eluent, and the mycotoxins were eluted from the antibodies.
Before use, restore the affinity column to room temperature, let off the protective solution of the column, and then carry out the process of sample loading, washing and elution.
- The sample solution on the column should be neutral in acid and alkali, without suspended matter, and the methanol concentration should not be too high
- When the sample is a complex sample (such as high protein, high oil, acidic samples), the extract can be extracted twice to improve the extraction effect.
- The speed of sample passing through the column and methanol elution should be slow and uniform, about 1-2 drops per second
- The optimal temperature for the antigen-antibody reaction is around 25°C. The ambient temperature required for the experiment is preferably room temperature 24°C-28°C. Too high or too low temperature will have a certain impact on the recovery rate of the affinity column. . The affinity column needs to be returned to room temperature before experimenting
- The affinity column should not be in an anhydrous state, the agar is easy to dry and shrink, which will affect the quality of the affinity column.
- Affinity column storage conditions are 4℃-8℃. Remember that the low temperature below 0°C will seriously affect the physical structure of the affinity column and the quality of the affinity column because the buffer freezes.
- Methanol is generally selected as the reagent for elution. To enhance the elution effect, the volume of methanol can be increased.
- After the sample liquid is purified by immunoaffinity column, it is better to pass the eluate through a 0.45μm or smaller pore size filter.