Files in this item

FilesDescriptionFormat

application/pdf

application/pdf9702595.pdf (8MB)Restricted to U of Illinois
(no description provided)PDF

Description

Title:Properties of immunoglobulin Fv domains: Influence of valence and protein dynamics on antibody/antigen interactions
Author(s):Mallender, William David
Doctoral Committee Chair(s):Voss, Edward W., Jr.
Department / Program:Microbiology
Discipline:Microbiology
Degree Granting Institution:University of Illinois at Urbana-Champaign
Degree:Ph.D.
Genre:Dissertation
Subject(s):Biology, Molecular
Chemistry, Biochemistry
Health Sciences, Immunology
Abstract:Anti-metatype antibodies are immunoglobulins specific for liganded antibody active sites that, upon binding, delay dissociation of primary ligand resulting in augmented primary antibody affinity for ligand. Anti-metatype antibodies, both monoclonal and polyclonal, specific for the 4-4-20/fluorescein complex, have been generated and extensively studied. Preliminary studies identified reduced variable domain dynamics as the reason for 4-4-20 affinity for fluorescein enhancement due to anti-metatype antibody binding. Differences in activity levels between monoclonal and polyclonal anti-metatype reagents, however, implicated valence, specificity and affinity for generating a functional anti-metatype stabilization effect. In order to investigate this hypothesis, a bivalent-bispecific single-chain antibody (BiSCA), consisting of the SCA derivatives of IgG 4-4-20 and IgG 04-01 (anti-ssDNA), was constructed and characterized as a prelude to construct novel BiSCA anti-metatype proteins. Following primary structure determination and failure to successfully produce active anti-metatype SCA molecules from candidate monoclonal antibodies, a bispecific anti-metatype monoclonal antibody was produced by chemical cross-linking methods. Results from studies comparing the activity of various anti-metatype antibody reagents confirmed the roles of valence and multispecificity in the anti-metatype stabilization effect. In addition, further analyses of the effect of anti-metatype stabilization on antibody variable domains was accomplished by construction and characterization of the Fv derivative of 4-4-20. This active site derivative protein, unlike SCA, lacked the interdomain linker peptide, permitting examination of the effect of anti-metatype stabilization on variable domain association. Such studies yielded significant information concerning the dependence of antibody active site dynamics on high affinity antigen binding and the importance of antibody constant domains on the maintenance of these dynamics. Collectively, studies of this nature could be applied to other protein systems where multivalent protein subunit associations are employed for effective ligand binding.
Issue Date:1996
Type:Text
Language:English
URI:http://hdl.handle.net/2142/20562
ISBN:9780591088663
Rights Information:Copyright 1996 Mallender, William David
Date Available in IDEALS:2011-05-07
Identifier in Online Catalog:AAI9702595
OCLC Identifier:(UMI)AAI9702595


This item appears in the following Collection(s)

Item Statistics