Ying Zheng

Defense Date

10-14-2010

Graduation Date

Fall 2010

Availability

Immediate Access

Submission Type

dissertation

Degree Name

PhD

Department

Pharmaceutics

School

School of Pharmacy

Committee Chair

Wilson Meng

Committee Member

James Drennen

Committee Member

Philip Auron

Committee Member

Ellen Gawalt

Committee Member

Peter Wildfong

Keywords

EAK, Histidine, Insoluble structure, Monoclonal antibody, Regulatory T cells, Self-assembling peptides

Abstract

This study investigated a strategy by which antibodies were displayed on a gel-like substance to engage T cells. The substance, a peptidic composite, was characterized in vitro and explored as an injectable system in vivo. The composite consists of two amphiphilic peptides, AEAEAKAKAEAEAKAK (referred to as "EAK") and AEAEAKAKAEAEAKAKHHHHHH ("EAKH6"). Spectroscopic analysis showed the two peptides integrated into a single structure. Prior to combination, conformational analysis revealed that EAKH6 adopts a mixed alpha-helix/bata-strand conformation. In the presence of EAK, EAKH6 exists predominantly in a beta;-strand conformation. Using nickel-bound horseradish peroxidase as a probe, the composite of EAK-EAKH6 was found to display His-tags. T-cell-specific antibodies were found stably displayed on the EAK-EAKH6 assembly using recombinant protein A/G and anti-hexahistidine antibody as an adaptor. When mounted with an anti-CD4 antibody, the system was shown to capture CD4 T cells in a mixed population of lymphocytes. Antibodies were concentrated in the subcutaneous space in mice when co-administered with EAK and EAKH6 along with protein A/G and anti-hexahistidine antibody as an aqueous (deionized water)

Format

PDF

Language

English

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