Technology of compact MAb and its application for medicinal plant breeding named as missile type molecular breeding.

Single chain fragment-variable (scFv) enhanced solasodine glycoside accumulation in Solanum khasianum hairy root cultures transformed by the ScFv solamargine (As)-scFv gene. The scFv protein was expressed at a high level in inclusion bodies of E. coli. After being renatured, the scFv protein was purified in a one-step manner by metal chelate affinity chromatography. The yield of refolded and purified scFv was 12.5 mg per 100 ml of cell culture. The characteristics of the As-scFv expressed in E. coli and transgenic hairy roots were similar to those of the parent monoclonal antibody (MAb). The expression of scFv protein provides a low cost and a high yield of functional scFv antibody against solamargine. The full linear range of the ELISA assay using scFv was extended from 1.5-10 µg/ml. The expressed anti-solamargine scFv protein could be useful for determination of total solasodine glycoside content in plant samples by ELISA. Solasodine glycoside levels in the transgenic hairy root were 2.3-fold higher than that in the wild-type hairy root based on the soluble protein level and binding activities. The As-scFv expressed in S. khasianum hairy roots enhanced solasodine glycosides accumulation and provide a novel medicinal plant breeding methodology that can produce a high yield of secondary metabolites.

Anti-solasodine glycoside single-chain Fv antibody stimulates biosynthesis of solasodine glycoside in plants.

We constructed a recombinant antibody fragment--single chain fragment-variable (scFv) antibody--derived from hybridoma cell lines to control the concentration of solasodine glycosides in hairy root cultures of Solanum khasianum transformed by the anti-solamargine (As)-scFv gene. The properties of the As-scFv protein expressed in Escherichia coli were almost identical to those of the parent monoclonal antibody (MAb). Up to 220 ng recombinant As-scFv was expressed per milligram of soluble protein in transgenic hairy root cultures of S. khasianum. The concentration of solasodine glycosides was 2.3-fold higher in the transgenic than in the wild-type hairy root, as reflected by the soluble As-scFv level and antigen binding activities. These results suggested that the scFv antibody expressed in transgenic hairy roots controlled the antigen level, thus representing a novel plant breeding methodology that can produce secondary metabolites.

[Sensitization to paprika: anaphylaxis after intake and rhinoconjunctivitis after contact through airways]. / Sensibilización al pimentón: anafilaxia por ingestión y rinoconjuntivitis tras contacto por vía aérea.

naphylaxis after the intake of paprika and with rhinoconjunctivitis symptoms related to her current occupation, as spices-and-condiment seller, induced the patient to attend to a medical consult. The patient tolerated another solanaceous in her diet. The study was based on skin test using own extracts of different solanaceous, both at room temperature and after warming them up. Skin test with prick-by-prick test were also carried out with fresh solanaceous. All skin tests showed a positive result. The study was completed with conjunctival challenge-test using all the different extracts (positive result with paprika) and use-test with fresh solanaceous (positive result with paprika). In laboratory study, specific IgE was detected to all the solanaceous, as well as histamine release-test positive. Crossed-reactivity between the paprika antigenic determinant recognized by the patient and the one presented by the rest of solanaceous could not be detected using RAST-inhibition studies. This results and the good tolerance of other solanaceous in diet could show that the paprika's antigenic determinant, what induced allergic symptoms in the patient, was not present in the rest of solanaceous.