Carrier protein is a kind of macromoleculecan trigger an immune responseitself, many different kinds of carrier protein couplingwithsmall molecules can prepare complete antigen.Common carrier including bovineserum albumin, keyhole hemocyanin, ovalbumin, and cattle thyroglobulin etc.
For the choice of carrier protein, mainlyrelated to its solubility, size, coupling group, immunogenicity, costprice andother factors.Currently the most widely used in marketis bovine serum albuminBSA (3D structure in Fig. 1), which mainly due to the BSA have chemicalstability, not easy denaturation, molecule contains a variety of amino, andhigh contentlysine,cheap and easy to get.But as BSA in some immune detectionexperiments act as a blocker to restrict its usability.The second morechoicesarekeyhole hemocyanin KLH and ovalbumin OVA.
Briefly introduce the method of hapten andcarrier coupling:
1, with carboxyl or amino compounds, usingtwo carbide imide method and mixed anhydride method;
2, containing sulphur compounds, usingmaleic imide method;
3, the molecular structure of compoundswith amino class, glutaraldehyde method, one of the most commonly used method.
General hapten and carrier protein,according to above method, choosing the appropriate coupling ratio at roomtemperature or 4 ℃ reaction for one night, then with a separation column or dialysisbag 2 d, achieve fully antigen.
For fully antigen, can use uv scanningmethod, comparing the change of ultraviolet absorption with both couplingproductand un-coupling carrier protein.Artificial fully antigen ultraviolet absorptioncharacteristics contain or have different from the original hapten and carrierprotein, can ensure coupling successful, then through operation, get thecoupling rate.
Biobool after a great deal of research anddevelopment, can provide a variety of types coupling small molecular compoundsand polypeptide, for researchers choose to use.
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