Expression and purification of DT-EGF fusion protein in Pichia pastoris for preclinical studies : a thesis /
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| Format: | Thesis Book |
| Language: | English |
| Published: |
[College Station, Tex.] :
[Texas A&M University System Health Science Center],
[2009]
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| Subjects: |
| Abstract: | ABSTRACT: EGF is a growth factor that plays an important role in cell proliferation. Upon binding to its receptor EGFR, a transmembrane glycoprotein, a series of signaling cascades will be initiated which ultimately regulates cell growth and proliferation. Studies have shown that overexpression and upregulation of EGFR can be found in a wide spectrum of human cancer. Currently available therapies are generally not curative and are often accompanied by local and systemic toxicities. With no improvement in clinical outcome, a more effective and less toxic therapy is highly desired. Here, we report to be the first to synthesize recombinant DT-EGF fusion protein in yeast using Pichia pastoris as the expression system. We hae successfully constructed a gene expression cassette to express DT-EGF, synthesized and purified preclinical grade DT-EGF, and characterized purified DT-EGF. Purified DT-EGF molecular weight was determined to be 48.4 KDa by mass spectrometry and its average potency against EGFR1 cells was 5.97 pM. Final purity of the purified protein was greater than 98% with <0.04% protein aggregation. |
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| Item Description: | Vita. "Major Subject: Medical Sciences". "Submitted to the Office of Research and Graduate Studies of The Texas A&M University System Health Science Center in partial fulfillment for the requirements for the degree of Master of Science August 2009." Approved as to style and content by: Arthur E. Frankel, Jung Hee Woo, Cynthia Meininger. |
| Physical Description: | xi, 69 leaves : illustrations ; 28 cm. |
| Bibliography: | Includes bibliographical references (leaves 63-68). |