High-affinity rat anti-fluorescein monoclonal antibody with unique fine specificity properties including differential recognition of dynamic ligand analogues

Journal of Molecular Recognition : JMR
S D MiklaszE W Voss

Abstract

The ability of antibodies to specifically select and stabilize through binding one or more isomers of highly dynamic ligands remains a relatively unexplored immunochemical problem. The experimental strategy employed in this study was to elicit homogeneous antibodies to polyaromatic fluorescein which exists in one isomeric form. The binding properties of a monoclonal rat antifluorescein antibody specific to a given isomer were quantitatively studied to determine the capacity to bind dynamic analogues of fluorescein which exists in multiple isomers. To generate monoclonal anti-fluorescein antibodies that reacted with specific dynamic analogues of fluorescein possessing unconjugated aromatic ring systems, immune spleenocytes from Lou/M rats immunized with FITC(I)-KLH were fused with Balb/c SP2/0-Ag14 murine myeloma cells forming rat-mouse hybridomas. Cell line P2A12-1-C8 was selected for further characterization from the original 23 stable rat hybrids, since it produced a monoclonal antibody with a binding affinity 2.0 x 10(10)/M for fluorescein based on dissociation rate measurements. P2A12-1-C8 exhibited significant reactivity with HPF and phenol red, which are dynamic structural analogues of the homologous fluorescein ligand. N...Continue Reading

References

Jan 1, 1977·Advances in Experimental Medicine and Biology·E W Voss, R M Watt
Mar 11, 1991·Biophysical Journal·C A Swindlehurst, E W Voss
Aug 1, 1989·Molecular Immunology·M A Dombrink-KurtzmanE W Voss
Dec 1, 1988·Molecular Immunology·M A Dombrink-Kurtzman, E W Voss
Jan 1, 1988·Annual Review of Biophysics and Biophysical Chemistry·H FrauenfelderR D Young
Nov 1, 1983·Journal of Biochemical and Biophysical Methods·J N Herron, E W Voss
Dec 30, 1982·Journal of Immunological Methods·S A NoemanT J Gill
Feb 1, 1983·Applied and Environmental Microbiology·R A Smith, J T Ulrich

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