Cryoprecipitation Properties and Structural Analysis of a High Affinity Anti-Fluorescein Immunoglobulin-M Antibody

A high affinity (K$\sb{\rm a} = 2.9 \times 10\sp $ M$\sp{-1}$) murine monoclonal anti-fluorescein IgM antibody (18-2-3) exhibiting low temperature insolubility in the absence of bound ligand has served as a model to study cryoprecipitation. Insolubility of 18-2-3 at low temperature (4$\sp\circ$C) had been shown to be reversible at higher temperatures and in the presence of fluorescyl ligand, indicating antigen binding site involvement. The primary objectives were to isolate and identify structural component(s) responsible for insolubility at low temperature. Procedures were developed for production and isolation of the monomeric subunit (IgM$\sb{\rm s}$) and Fab and (Fc)$\sb5$ fragments. Electrostatic interactions were implicated as being responsible for atypical low temperature insolubility of 18-2-3 since self-aggregation was sensitive to pH, ionic strength, temperature and protein concentration. Results indicated that interactions involving 18-2-3 antibody-combining sites with interactive sites in the Fc region of homologous IgM were responsible for the phenomenon of cryoprecipitation.

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

2014

Author dc:creator

• Dombrink-Kurtzman, Mary Ann

Contributors dc:contributor

• Voss, Edward W., Jr.

• Biology, Microbiology
• Health Sciences, Immunology

Identifier

(UMI)AAI8908666

OAI identifier oai:identifier

oai:www.ideals.illinois.edu:2142/71193