Prodrug Therapy for Breast Cancer Targeted by Single-Chain Antibodies F19 and 3S193 PDF Download
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Languages : en
Pages : 52
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Book Description
In Antibody-Directed Enzyme-Prodrug Therapy (ADEPT), antibody - enzyme constructs localize to tumor tissue the toxification of non-toxic prodrugs. Recombinant fusion proteins are expected to overcome the limitations of chemical conjugates. We have constructed fusion proteins of antibodies against the tumor antigens A33, Lewis-Y and fibroblast associated protein (PAP) with cytosine deaminase (CD), which converts 5-fluorocytosine (5-PC) into cytotoxic 5-fluorouracil (5-PU) - Initially using a T7 polymerase-based expression system, plasmid vectors had been designed for the cloning and expression of the fusion constructs in bacterial and Pichia pastoris expression systems. After initial production in E. coli, fusion proteins were produced in P. pastoris and purified from media supernatant with higher yield than from the previously used bacterial expression system. During the reporting period, work on this project was limited due to the ongoing transfer of funds from to the primary investigator's new institution in Germany. However, the expression systems used could be further optimized to achieve good fusion construct yields with pilot proteins. With funds now available, the project is well set to continue successfully with the planned animal experiments in the revised time frame.
Author:
Publisher:
ISBN:
Category :
Languages : en
Pages : 52
Get Book Here
Book Description
In Antibody-Directed Enzyme-Prodrug Therapy (ADEPT), antibody - enzyme constructs localize to tumor tissue the toxification of non-toxic prodrugs. Recombinant fusion proteins are expected to overcome the limitations of chemical conjugates. We have constructed fusion proteins of antibodies against the tumor antigens A33, Lewis-Y and fibroblast associated protein (PAP) with cytosine deaminase (CD), which converts 5-fluorocytosine (5-PC) into cytotoxic 5-fluorouracil (5-PU) - Initially using a T7 polymerase-based expression system, plasmid vectors had been designed for the cloning and expression of the fusion constructs in bacterial and Pichia pastoris expression systems. After initial production in E. coli, fusion proteins were produced in P. pastoris and purified from media supernatant with higher yield than from the previously used bacterial expression system. During the reporting period, work on this project was limited due to the ongoing transfer of funds from to the primary investigator's new institution in Germany. However, the expression systems used could be further optimized to achieve good fusion construct yields with pilot proteins. With funds now available, the project is well set to continue successfully with the planned animal experiments in the revised time frame.
