A recombinant lipid transfer protein from Cannabis sativa: IgE-binding properties in patients with symptoms to Cannabis

Methods To identify the mature peptide sequence of the lipid transfer protein (LTP) and to study the IgE-binding reactivity of a recombinant Can s 3 (rCan s 3), a cDNA was synthesized from total RNA of Cannabis sativa L. ssp. sativa cv. Kompolti leaves obtained from the botanical garden of the University Bonn. The LTP gene was amplified with a primer mix deduced from published amino acid sequences. The LTP variant was identified in five independent clones by sequencing in the pDrive vector system followed by the expression of the mature LTP in the pMAL-vector. After isolation of a soluble recombinant maltose-binding protein (MBP)-Can s 3 fusion protein, aliquots were biotinylated and coupled to Streptavidin-ImmunoCAPs.