Recombinant allergens and antibodies are important for diagnostic,
therapeutic, food processing and quality verification purposes. Thus,
researchers from VTT Technical Research Centre of Finland and International
Centre for Genetic Engineering and Biotechnology developed a barley-based
production system for detection of bovine milk allergen �-lactoglobulin
(BLG) specific immunoglobulin E antibody (D1 scFv).

The researchers found that the expression level of the antibody in the best
barley cell clone was 0.8-1.2 mg/kg fresh weight, which remained constant
over an expression period of 3 weeks. In the case of barley grains, the
highest stable productivity (followed up to T2 grains) was obtained when the
D1 scFv cDNA was expressed under a seed-specific Glutelin promoter.

Translational fusion of ER retention signal significantly enhanced the build
up of recombinant antibody. Furthermore, lines without ER retention signal
lost D1 scFv accumulation in T2 grains. Pilot scale purification resulted in
a yield of 0.47 mg of D1 scFv (31 kD) with high purity. Further analysis
revealed that 29 percent of the purified protein was completely functional.
The results indicate that the developed barley-based expression system can
be used for application in the processing of dairy milk products as well as
in detecting allergens from foods possibly contaminated by
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O9bf054af&e?99440571> bovine milk.

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