Tehran University of Medical Sciences
Office of Vice-Chancellor for Global Strategies & International Affairs
International Human Capacity Development (IHCD)
Code : 9822-349940      Publish Date : Saturday, June 20, 2015 Visit : 2559

Intl. Congress form | International Congress Report | International Congress Report For Students and Staff | 8th Conference on Recombinant Protein Production – RPP8

8th Conference on Recombinant Protein Production – RPP8
The report of 8th Conference on Recombinant Protein Production – RPP8 by Seyedeh Hoda Jazayeri
 
Application Code :
306-0115-0058
 
Created Date : Saturday, April 18, 2015 12:25:10Update Date : Tuesday, May 19, 2015 12:00:12
IP Address : 217.146.218.171Submit Date : Tuesday, May 19, 2015 12:00:39Email : h.jazayeri@yahoo.com
Personal Information
Name : Seyedeh Hoda
Surname : Jazayeri
School/Research center : School of Advanced Technology in Medicine
If you choose other, please name your Research center :  
Position : student
Tel : +98-21-22959626
Information of Congress
Title of the Congress : 8th Conference on Recombinant Protein Production – RPP8.
Title of your Abstract : Production of Recombinant Human Follicle Stimulating Hormone (rhFSH) in Chinese Hamster Ovary (CHO) Cell Line Using Fusion of the DHFR/MTX and IR/MAR Gene Amplification Methods
country : Spain
From : Wednesday, April 22, 2015
To : Friday, April 24, 2015
Abstract(Please copy/paste the abstract send to the congress) :
Abstract:
Background: Follicle stimulating hormone (FSH) is an effective gonadotropin used in infertility treatment. Production of Recombinant Human FSH (rhFSH) has been attempted by methotrexate (MTX) induction of inserted gene, located beside the dihydrofolate reductase (DHFR) locus. However, inefficiency disadvantages of this method have been reported, so far. 
Objectives: Here, we present a DHFR/MTX fusion method that applies a DNA fragment containing mammalian replication initiation region (IR) and matrix attachment region (MAR). 
Methods: The ORF regions of alpha and beta subunits of hFSH gene were cloned in a shuttle vector and used for co-transfection of CHO-DHFR‾ cells along with the “IR/MAR gene amplification reagent. Then, co-selection of clones using MTX and blasticidin S antibiotics, followed by a precise drugs increasing dosage profile to stimulate gene amplification have been performed. The expression of gene construct entailing rhFSH in CHO-DHFR‾ cell line was verified by SDS PAGE and western blot. The construct copy number as well as mRNA or protein expression levels were determined by real time-PCR and ELISA analyses, respectively.
Results: Stable CHO-DHFR‾ clones with significantly higher rhFSH expression level were established. Enhanced amplification of the copy number of a co-transfected expression vector was obtained, leading to high rhFSH protein production efficacy. 
Conclusion: Collectively, our data shows IR/MAR-Dhfr fusion, together with MTX and blasticidin S co-selection, is a reliable methodology for isolation, and high-yield production of rhFSH, as compare to the sole application of either the DHFR/MTX or IR/ MAR methods.
Keywords of your Abstract : Keywords: Recombinant Follicle stimulating hormone, CHO-DHFR‾, DHFR, IR/MAR.
Acceptance Letter : http://gsia.tums.ac.ir/images/UserFiles/24518/Forms/306/poster acceptance_3.pdf
The presentation : Poster
The Cover of Abstract book : http://gsia.tums.ac.ir/images/UserFiles/24518/Forms/306/cover_page.pdf
Published abstract in the abstract book with the related code : http://gsia.tums.ac.ir/images/UserFiles/24518/Forms/306/abstracct.pdf
Where has your abstract been indexed? : none
If you choose other, please name :  
The Congress Reporting Form
How many volunteers were present at the Congress? : about 300 people
Delegates from which countries presented in the congress? : Spain, Austria, Germany, U.k., U.S.A, Netherland, etc
Were the delegates of any other organizations present in the congress? : Yes
If yes, please write the names of the organizations in the box : BOKU, LONZA, NOVOZYMES
What were the responses to your talking points? Were specific questions or concerns raised? : There were some question about the effect and the mechanism of IR/MAR on protein production and expression. one PhD student advised me for using signal peptide in construct for having better expression
If you met staff members, please list their full names & positions. : -
Please inform us if there are any follow up actions we need to talk with the members of the congress : As I was attended in the conference with my professor, Dr Mohammad Hossein Sanati from national institute of genetic engineering and biotechnology and and also one of my friends on behalf of Royan institute the collaborations will continue from these two institute. There were some scholarships and grants from companies and universities for PhD students
Your experiences about the travel processes(Providing ticket, accommodation,...) : Travel expenses was provided by myself and hotel was booked by congress but there was some problems that made it unpleasant because the time of our arrival time and the hotel reservation was different which made some mistakes for us.
Please give a briefing of your own observations and outcomes of the congress: : Scientifically it was a useful conference specially it was focused on my field of expertise. It was a good opportunity to share ideas and become familiar with new strategies and techniques. According to the expression systems that can be used for protein production and the reality that protein expression in mammalian cells is more efficient specially because of the folding and glycosylation of proteins in this system but there are some disadvantages including low yield and high cost of these systems some alternative will be present in the conference such as prokaryotes with folding and glycosylation systems