Targeting cell-surface tyrosine kinase receptors with recombinant antibody-like molecules / Norman Carl Peterson.

Format:

Book

Manuscript

Microformat

Thesis/Dissertation

Author/Creator:

Peterson, Norman Carl.

Contributor:

Greene, Mark I., advisor.

University of Pennsylvania.

Language:

English

Subjects (All):

Penn dissertations--Comparative Medicine.

Comparative Medicine--Penn dissertations.

Comparative Medicine.

Academic Dissertations as Topic.

Medical Subjects:

Comparative Medicine.

Academic Dissertations as Topic.

Local Subjects:

Penn dissertations--Comparative Medicine.

Comparative Medicine--Penn dissertations.

Physical Description:

viii, 117 pages : color illustrations ; 29 cm

Production:

1997.

Summary:

Homomeric and heteromeric interactions among cell-surface tyrosine kinase receptors belonging to the ErbB family lead to intracellular signaling cascades which are involved in cell activation, cytoskeletal interactions, and cellular transformation leading to neoplasia. Breast and ovarian cancer patients which over-express either p185$\sp{\rm c-erbB2}$ or epidermal growth factor receptor (EGFR) have a poorer prognosis than patients with tumors lacking over-expression of these receptors. Additionally, clinical and biochemical evidence suggests that heterodimerization of p185$\sp{\rm neu}$ and EGFR results in a potentiated growth signal making dual receptor over-expressing tumors even more aggressive. Monoclonal antibodies which specifically bind to p185$\sp{\rm neu}$ or EGFR, such as 7.16.4 and 225, respectively, can elicit tumor growth inhibitory effects upon transformed cells which over-express either or both of these receptors. In order to better understand these receptor-receptor and receptor-antibody interactions and to gain insights which may be useful in the production and design of an antibody-based anti-cancer therapeutic, novel small recombinant 7.16.4 and 225 single chain Fv fragments (scFv) were constructed, expressed, and characterized. Results presented here demonstrate that these recombinant antibody fragments can be produced and purified from bacterial cell lysates and retain binding activity. I also demonstrate that fusion of a 61 amino acid dimerization domain to 7.16.4 and 225 scFv (7.16.4dhlx and 225 dhlx) is sufficient to restore biological activity to these recombinant proteins. However, improper protein folding and aggregation limit the ability of 7.16.4 dhlx and 225 dhlx to form functional heterodimeric complexes capable of simultaneously binding p185$\sp{\rm neu}$ and EGFR.

Notes:

Supervisor: Mark I. Greene.

Thesis (Ph.D. in Comparative Medicine)--Graduate School of Arts and Sciences, University of Pennsylvania, 1997.

Includes bibliographical references.

Local Notes:

University Microfilms order no.: 98-14901.

OCLC:

187457388