Technology of Compact Monoclonal Antibody and its Application for Medicinal Plant Breeding Named as Missile Type Molecular Breeding.
Summary of "Technology of Compact Monoclonal Antibody and its Application for Medicinal Plant Breeding Named as Missile Type Molecular Breeding."
Single chain fragment-variable (scFv) antibody enhanced solasodine glycoside accumulation in Solanum khasianum hairy root cultures transformed by the anti-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 protein expressed in E. coli and transgenic hairy roots were almost identical 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 As-scFv level and antigen binding activities. The As-scFv expressed in transgenic hairy roots enhanced the antigen accumulation and provide a novel medicinal plant breeding methodology that can produce a high yield of secondary metabolites.
Affiliation
Khon Kaen University, Khon Kaen, 40002, Thailand. waraporn@kku.ac.th.
Journal Details
This article was published in the following journal.
Name: Current drug discovery technologies
ISSN: 1875-6220
Pages:
Links
- PubMed Source: http://www.ncbi.nlm.nih.gov/pubmed/21143130
- DOI: http://dx.doi.org/
Medical and Biotech [MESH] Definitions
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Various mixtures of fats, waxes, animal and plant oils and solid and liquid hydrocarbons; vehicles for medicinal substances intended for external application; there are four classes: hydrocarbon base, absorption base, water-removable base and water-soluble base; several are also emollients.
Rosmarinus
A plant genus of the LAMIACEAE family. It is known as a spice and medicinal plant.
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