Derivation and Properties of Recombinant Fab Antibodies to the Phenylurea Herbicide Diuron Academic Article uri icon

abstract

  • The Fab domain sequences from messenger RNA of a mouse hybridoma cell line secreting a monoclonal antibody (MAb) specific for the phenylurea herbicide diuron were amplified and inserted into the M13 phagemid vector pComb8. Phage displaying diuron-specific Fab fragments were selected by binding to magnetic beads coated with diuron hapten conjugates, and eluted with diuron. Soluble rFab in E. coli lysates bound diuron with half-maximal inhibition (I50) of 1.6-12 ppb (ng/ mL) in indirect and direct competition enzyme immunoassays (EIAs). The optimal competing hapten was different for indirect EIAs with proteolytic or recombinant Fabs and the intact MAb. Selectivity of rFabs for phenylureas was similar to that of the original MAb and its proteolytic Fab fragment and was not significantly affected by exchange of heavy and light chains among rFab clones, indicating that a single antibody species had been cloned.

published proceedings

  • Journal of Agricultural and Food Chemistry

author list (cited authors)

  • Scholthof, K., Zhang, G., & Karu, A. E.

citation count

  • 15

complete list of authors

  • Scholthof, Karen-Beth G||Zhang, Guisheng||Karu, Alexander E

publication date

  • April 1997