Recombinant single-chain antibodies against s-triazines

Recombinant antibody (RAb) technology is expected to play all essential role in future food and environmental immunoanalysis. The RAb approach is not yet, however, considered as a standard method, mainly due to technical reasons. In order to establish a feasible protocol for the production of hapten-specific RAbs, the essential steps of the procedure are considered in detail, especially the polymerase chain reaction amplification of antibody coding genes and the utilization of immunomagnetic selection of phages displaying selective RAbs. The synthesis of functional recombinant single-chain Fv against s-triazine herbicides is used as an example. Expressed RAbs are characterized by immunoassay. Although RAb-based calibration curves were shifted toward higher concentrations by one order of magnitude, the cross-reactivity pattern of RAbs corresponded to the original monoclonal antibodies. Sequences of different RAb clones derived from the same hybridoma line only differed in a single silent mutation. This confirms the reliability of the RAb procedure.