Expression and Characterization of a Functional Single-Chain Variable Fragment (scFv) Protein Recognizing MCF7 Breast Cancer Cells in E. coli Cytoplasm

Allan AL, Vantyghem SA, Tuck AB, Chambers AF (2006) Tumor dormancy and cancer stem cells: implications for the biology and treatment of breast cancer metastasis. Breast Dis 26:87–98 Azemar M, Schmidt F (2000) Recombinant antibody toxins specific for erbB-2 and EGF receptor inhibit the in vitro growth of human head and neck cancer cells and cause rapid tumor regression in vivo. Int J Cancer 86:269–275 Bellinzoni M, De Rossi E, Branzoni M, Milano A, Peverali FA, Rizzi M, Riccardi G (2002) Heterologous expression, purification, and enzymatic activity of Mycobacterium tuberculosis NAD synthetase. Protein Expr Purif 25:547–557 Bessette PH, Aslund F, Beckwith J, Georgiou G (1999) Efficient folding of proteins with multiple disulfide bonds in the Escherichia coli cytoplasm. Proc Natl Acad Sci USA 96:13703–13708 Bishia WR, Rappuoli R, Murphy JR (1987) High-level expression of a proteolytically sensitive diphtheria toxin fragment in Escherichia coli. J Bacteriol 169:5140–5151 Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantitites of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254 Burton DR, Barbas CF (1994) Human antibodies from combinatorial libraries. Adv Immunol 57:191–280 Chae JS, Choi JK, Lim HT, Cha SH (2001) Generation of a murine single chain Fv (scFv) antibody specific for cucumber mosaic virus (CMV) using a phage display library. Mol Cells 1:12–17 Cummins E, Luxenberg DP, McAleese F, Widom A, Fennell BJ, Darmanin-Sheehan A, Whitters MJ, Bloom L, Gill D, Cunningham O (2008) A simple high-throughput purification method for hit identification in protein screening. J Immunol Methods 339:38–46 Dutta S, Ware LA, Barbosa A, Ockenhouse CF, Lanar DE (2001) Purification, characterization, and immunogenicity of a disulfide cross-linked Plasmodium vivax vaccine candidate antigen, merozoite surface protein 1, expressed in Escherichia coli. Infect Immun 69:5464–5470 Dutta S, Lalitha PV, Ware LA, Barbosa A, Moch JK, Vassel MA, Fileta BB, Kitov S, Kolodny N, Heppner DG, Haynes JD, Lanar DE (2002) Purification, characterization, and immunogenicity of the refolded ectodomain of the plasmodium falciparum apical membrane antigen 1 expressed in Escherichia coli. Infect Immun 70:3101–3110 Freund C, Ross A, Pluckthun A, Holak TA (1994) Structural and dynamic properties of the Fv fragment and the single-chain Fv fragment of an antibody in solution investigated by hetero-nuclear three-dimensional NMR spectroscopy. Biochemistry 33:3296–3303 Gattenlohner S, Jorissen HJ, Huhn M, Vincent A, Beeson D, Tzartos S, Mamalaki A, Etschmann B, Muller-Hermelink HK, Koscielniak E, Barth S, Marx A (2010) A human recombinant autoantibody-based immunotoxin specific for the fetal acetylcholine receptor inhibits rhabdomyo-sarcoma growth in vitro and in a murine transplantation model. J Biomed Biotech 11:10–21 Hu X, O’Hara L, White S, Magner E, Kane M, Wall JG (2007) Optimisation of production of a domoic acid-binding scFv antibody fragment in Escherichia coli using molecular chaperones and functional immobilisation on a mesoporous silicate support. Protein Expr Purif 52:194–201 Kaomek M, Mizuno K, Fujimura T, Sriyotha P, Cairns JR (2003) Cloning, expression, and characterization of an antifungal chitinase from Leucaena leucocephala de Wit. Biosci Biotechnol Biochem 67:667–676 Kashmiri SVS (2001) Multivalent single-chain antibodies for radioimaging of tumors. J Nucl Med 42:1528–1529 Krebber MA, Burmester J, Pluckthun A (1996) Inclusion of an upstream transcriptional terminator in phage display vectors abolishes background expression of toxic fusions with coat protein g3p. Gene 178:71–74 Lehmann K, Hoffmann S, Neudecker P, Suhr M, Becker WM, Rosch P (2003) High-yield expression in Escherichia coli, purification, and characterization of properly folded major peanut allergen Ara h 2. J Protein Expr Purif 31:250–259 Nilsson BL, Kiessling LL, Raines RT (2000) Staudinger ligation: a peptide from a thioester and azide. Org Lett (Am Chem Soc) 2:1939–1941 Pai LH, Batra JK, FitzGerald DJ, Willingham MC, Pastan I (1991) Anti-tumor activities of immunotoxins made of monoclonal antibody B3 and various forms of Pseudomonas exotoxin. Proc Natl Acad Sci USA 88:3358–3362 Peipp M, Kupers H, Saul D, Schlierf B, Greil J, Zunino SJ, Gramatzki M (2002) Recombinant CD7-specific single-chain immunotoxin is a potent inducer of apoptosis in acute leukemic T cells. Cancer Res 62:2848–2855 Prete SP, Cappelletti D, Baier S, Nasuti P, Guadagni F, De Vecchis L, Greiner JW, Bonmassar E, Graziani G, Aquino A (2002) Pharmacological modulation of carcinoembryonic antigen in human cancer cells: studies with staurosporine. Int Immunopharmacol 2:641–665 Reist CJ, Batra SW, Pegram CN, Bigner DD, Zalutsky MR (1997) In vitro and in vivo behavior of radiolabeled chimeric anti-EGFRvIII monoclonal antibody: comparison with its murine parent. Nucl Med Biol 24:637–647 Richard C, Drider D, Elmorjani K, Didier M, Prevost H (2004) Heterologous expression and purification of active divercin V41, a class IIa bacteriocin encoded by a synthetic gene in Escherichia coli. J Bacteriol 186:4276–4284 Scherf U, Benhar I, Webber KO, Pastan I, Brinkmann U (1996) Cytotoxic and antitumor activity of a recombinant tumor necrosis factor-B1(Fv) fusion protein on LeY antigen expressing human cancer cells. Clin Cancer Res 2:1523–1531 Shi PY, Kung WM, Chen JC, Yeh CH, Wang AHJ, Wang TF (2002) High throughput screening of soluble recombinant proteins. Protein Sci 111:714–1719 Singh R, Samant U, Hyland S, Pradip R, Chaudhari S, Winfried W, Bandyopadhyay D (2007) Target-specific cytotoxic activity of recombinant immunotoxin scFv (MUC1)-ETA on breast carcinoma cells and primary breast tumors. Mol Cancer Ther 6:72–89 Sorensen HP, Sperling-Petersen HU, Mortensen KK (2003) Production of recombinant thermo stable proteins expressed in Escherichia coli: completion of protein synthesis is the bottleneck. J Chromatogr B 786:207–214 Stewart EJ, Aslund F, Beckwith J (1998) Disulfide bond formation in the Escherichia coli cytoplasm: an in vivo role reversal for the thioredoxins. EMBO J 17:5543–5550 Tang CK, Gong XQ, Moscatello DK, Wong AJ, Lippman ME (2000) Epidermal growth factor receptor vIII enhances tumorigenicity in human breast cancer. Cancer Res 60:3081–3087 Villaverde A, Carrio MM (2003) Protein aggregation in recombinant bacterial biological role of inclusion bodies. Biotechnol Lett 25:1385–1395 Wikstrand CJ, McLendon RE, Friedman AH, Bigner DD (1997) Cell surface localization and density of the tumor-associated variant of the epidermal growth factor receptor, EGFRvIII. Cancer Res 57:4130–4140 Yuan LD, Hua ZC (2002) Expression, purification, and characterization of a biologically active bovine entero-kinase catalytic subunit in Escherichia coli. Protein Expr Purif 25:300–304 Zhou H, Liu Y, Cheung LH, Kim S, Zhang W, Mohamedali KA, Anand P, Hittelman WN, Aggarwal BB, Rosenblum MG (2010) Characterization and mechanistic studies of a novel melanoma-targeting construct containing IκBα for specific inhibition of nuclear factor-κB activity. Neoplasia 12:766–777