Kinetic Analysis of High Affinity Polyclonal Anti-Fluorescyl Immunoglobulin-G Antibodies

Mathematical and experimental tools for the measurement and assessment of association and dissociation kinetics in anti-fluorescyl antibodies were developed in these studies. An analytic procedure for the characterization of heterogeneous first order kinetics was derived. This procedure was based on Laplace transformation of the gamma distribution function, which yielded an exact rate law. Kinetic assays were developed for the measurement of both association and dissociation rates. Association rates were measured by following the increase in fluorescence quenching observed when anti-fluorescyl antibodies were added, in excess, to aqueous fluorescein. The reaction was made pseudo-first order to enable use of the same data processing routines for analysis of both association and dissociation kinetics. Dissociation rates were measured by following the increase in fluorescence intensity after the addition of excess 5-aminofluorescein to a population of fluorescyl-liganded antibodies. Since 5-aminofluorescein was a non-fluorescent competitive inhibitor of fluorescein, the increase in fluorescence intensity was directly attributable to the dissociation of fluorescyl-ligand.

Name thesis:degree_name

Ph.D.

Level thesis:degree_level

Dissertation

Discipline thesis:degree_discipline

Microbiology

Grantor

University of Illinois at Urbana-Champaign

Year dc:date

1981

Author dc:creator

• Herron, James Nelson

• Chemistry, Biochemistry

Language dc:language

eng

Identifier

(UMI)AAI8203489

OAI identifier oai:identifier

oai:www.ideals.illinois.edu:2142/68095