Next Article in Journal
Cholesterol-Lowering Effect of Beta Glucan Extracted from Saccharomyces cerevisiae in Rats
Previous Article in Journal
In Vitro Study on Antihypertensive and Antihypercholesterolemic Effects of a Curcumin Nanoemulsion
 
 
Scientia Pharmaceutica is published by MDPI from Volume 84 Issue 3 (2016). Previous articles were published by another publisher in Open Access under a CC-BY (or CC-BY-NC-ND) licence, and they are hosted by MDPI on mdpi.com as a courtesy and upon agreement with Austrian Pharmaceutical Society (Österreichische Pharmazeutische Gesellschaft, ÖPhG).
Order Article Reprints
Font Type:
Arial Georgia Verdana
Font Size:
Aa Aa Aa
Line Spacing:
Column Width:
Background:
Article

Construction and Periplasmic Expression of the Anti-EGFRvIII ScFv Antibody Gene in Escherichia coli

1
Research Center for Biotechnology, Indonesian Institute of Sciences (LIPI), Jalan Raya Bogor Km. 46, 16911, Cibinong, Bogor, Indonesia.
2
School of Pharmacy, Bandung Institute of Technology, Jalan Ganesha 10, 40132, Bandung, Indonesia.
*
Author to whom correspondence should be addressed.
Sci. Pharm. 2016, 84(1), 141-152; https://doi.org/10.3797/scipharm.ISP.2015.06
Received: 5 September 2015 / Accepted: 15 December 2015 / Published: 14 February 2016

Abstract

In the previous study, we constructed an expression vector carrying the anti- EGFRvIII scFv antibody gene with VH-linker-VL orientation. The proteins were successfully produced in the periplasmic space of Escherichia coli. In this study, we substituted the inserted DNA with VL-linker-VH orientation of the anti- EGFRvIII scFv gene and analyzed its expression in E. coli. The DNA fragment was amplified from its cloning vector (pTz-rscFv), subsequently cloned into a previous expression vector containing the pelB signal sequence and his-tag, and then transformed into E. coli TOP10. The recombinant plasmids were characterized by restriction, PCR, and DNA sequencing analyses. The new anti-EGFRvIII scFv antibody proteins have been successfully expressed in the periplasmic compartment of E. coli Nico21(DE3) using 0.1 mM final concentration of IPTG induction. Total proteins, soluble periplasmic and cytoplasmic proteins, solubilized inclusion bodies, and extracellular proteins were analyzed by SDS-PAGE and Western Blot analyses. The results showed that soluble scFv proteins were found in all fractions except from the cytoplasmic space.
Keywords: scFv; EGFRvIII; Periplasmic expression; PelB; E. coli scFv; EGFRvIII; Periplasmic expression; PelB; E. coli

Share and Cite

MDPI and ACS Style

DEWI, K.S.; RETNONINGRUM, D.S.; RIANI, C.; FUAD, A.M. Construction and Periplasmic Expression of the Anti-EGFRvIII ScFv Antibody Gene in Escherichia coli. Sci. Pharm. 2016, 84, 141-152. https://doi.org/10.3797/scipharm.ISP.2015.06

AMA Style

DEWI KS, RETNONINGRUM DS, RIANI C, FUAD AM. Construction and Periplasmic Expression of the Anti-EGFRvIII ScFv Antibody Gene in Escherichia coli. Scientia Pharmaceutica. 2016; 84(1):141-152. https://doi.org/10.3797/scipharm.ISP.2015.06

Chicago/Turabian Style

DEWI, Kartika Sari, Debbie Sofie RETNONINGRUM, Catur RIANI, and Asrul Muhamad FUAD. 2016. "Construction and Periplasmic Expression of the Anti-EGFRvIII ScFv Antibody Gene in Escherichia coli" Scientia Pharmaceutica 84, no. 1: 141-152. https://doi.org/10.3797/scipharm.ISP.2015.06

Article Metrics

Back to TopTop