Characterization of a monoclonal antibody specific to a flavonol 2'-O-glucosyltransferase.
A monoclonal antibody to a partially purified preparation of 2'-O-glucosyltransferase was produced by in vitro immunization of spleen cells from BALB/c mice, followed by fusion with mouse myeloma cells. Hybridoma culture supernatants were screened by enzyme-linked immunosorbent assay for (i) their ability to produce immunoglobulins and (ii) their immunoreactivity with a partially purified enzyme preparation. The majority of the immunoglobulin-producing hybridomas were IgM secretors. Two highly immunoreactive IgM-secreting clones were chosen for further characterization. The supernatant fraction from a culture of one of these clones displayed 50% inhibition of the 2'-O-glucosyltransferase activity. The native form of the 2'-O-glucosyltransferase was essential for recognition, suggesting that the epitope recognized by the antibody is a conformational discontiguous one.
Latchinian
L
L
Department of Biology, Concordia University, Montreal, Que., Canada.
Ibrahim
R K
RK
D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis*; D015153:Blotting, Western; D002459:Cell Fusion; D002478:Cells, Cultured; D055598:Chemical Phenomena; D002621:Chemistry; D004797:Enzyme-Linked Immunosorbent Assay; D005964:Glucosyltransferases; D006825:Hybridomas; D051379:Mice; D013154:Spleen / Q000276:immunology
D000911:Antibodies, Monoclonal; C060953:flavonol 2'-O-glucosyltransferase; D005964:Glucosyltransferases