Selection of beet necrotic yellow vein virus specific single-chain Fv antibodies from a semi-synthetic combinatorial antibody library

R.A. Griep, C. van Twisk, A. Schots

Research output: Contribution to journalArticleAcademicpeer-review

13 Citations (Scopus)

Abstract

Methods for the generation of monoclonal antibodies against plant viruses are limited because current hybridoma techniques do not allow efficient exploitation of the immune repertoire. Moreover, the immunization procedures often lead to a bias towards an immunodominant contaminant in the immunogen preparation and not to the plant virus itself. The selection of six different single-chain antibody variable fragments (scFv) against beet necrotic yellow vein virus from a semi-synthetic human combinatorial antibody library showed the feasibility of the phage display system. No bias towards minor contaminants in the purified virus preparation was observed in ELISA, as all the selected scFvs reacted only with beet necrotic yellow vein virus infected plant homogenates. In addition, two of the isolated beet necrotic yellow vein virus-specific scFvs could be produced in E. coli as a scFv fusion protein with alkaline phosphatase, and were applied in ELISA as specific ready to use antibody-enzyme conjugates. Because of their specificity, these antibodies have potential to be used as reagents in sensitive diagnostic assays for routine testing for beet necrotic yellow vein virus in sugar beets.
Original languageEnglish
Pages (from-to)147-156
JournalEuropean Journal of Plant Pathology
Volume105
DOIs
Publication statusPublished - 1999

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