Novel recombinant protein constructs for improved insulin-like growth factor-1 delivery

Four novel recombinant protein constructs were designed to improve the delivery of insulin-like growth factor-1 (IGF-1). The IGF-1 sequence was subcloned into a HisMax plasmid and expressed at low levels by transient transfection in 293 cells. Site-specific mutagenesis was used to insert all four construct sequences into the IGF-1 HisMax plasmid, and to insert a secretory sequence into all four construct plasmids. A stable cell line selection in 293 and CHO cells using Zeocin was attempted. The four construct sequences were then subcloned from the HisMax plasmids to TrcHis plasmids and expressed in bacteria for higher production efficiency. Ni-NTA purified protein was detected in three of the four constructs. These proteins may ultimately be useful for myocardial delivery of IGF-1 to facilitate cardiac repair.

Department dc:contributor.department

Massachusetts Institute of Technology. Dept. of Mechanical Engineering.

Grantor dc:publisher

Massachusetts Institute of Technology

Year dc:date.issued

2006

Author dc:creator

• Was, Adam (Adam S.)

Advisor dc:contributor.advisor

• Richard T. Lee.

• Mechanical Engineering.

Statement dc:rights

• M.I.T. theses are protected by copyright. They may be viewed from this source for any purpose, but reproduction or distribution in any format is prohibited without written permission. See provided URL for inquiries about permission.

Licence dc:rights.uri

• http://dspace.mit.edu/handle/1721.1/7582

Language dc:language.iso

eng

Handle dc:identifier.uri

http://hdl.handle.net/1721.1/36685

OAI identifier oai:identifier

oai:dspace.mit.edu:1721.1/36685