High yield expression of recombinant human proteins with the transient transfection of HEK293 cells in suspension

Journal of Visualized Experiments

Volumn 2015, Issue 106, 2015, Pages

  Subedi, Ganesh P ^a   Johnson, Roy W ^b   Moniz, Heather A ^b   Moremen, Kelley W ^b   Barb, Adam ^a  

^a IOWA STATE UNIVERSITY   (United States)

^b UNIVERSITY OF GEORGIA   (United States)

Author keywords

Cellular Biology; Fc; Glycoprotein; Human HEK293F and HEK293S cells; Immunoglobulin G; Issue 106; Receptor; Recombinant proteins

Indexed keywords

GLYCOPROTEIN; RECOMBINANT PROTEIN;

AFFINITY CHROMATOGRAPHY; ANIMAL; BIOSYNTHESIS; GENETIC TRANSFECTION; GENETICS; HEK293 CELL LINE; HUMAN; ISOLATION AND PURIFICATION; PROCEDURES; PROTEIN PROCESSING; RAT;

ANIMALS; CHROMATOGRAPHY, AFFINITY; GLYCOPROTEINS; HEK293 CELLS; HUMANS; PROTEIN PROCESSING, POST-TRANSLATIONAL; RATS; RECOMBINANT PROTEINS; TRANSFECTION;

EID: 84952836951     PISSN: 1940087X     EISSN: None     Source Type: Journal    
DOI: 10.3791/53568     Document Type: Article

References (52)

1. Terpe, K. Overview of bacterial expression systems for heterologous protein production: from molecular and biochemical fundamentals to commercial systems. Appl Microbiol Biotechnol. 72(2), 211-222, (2006).
2. Sorensen, H. P., & Mortensen, K. K. Advanced genetic strategies for recombinant protein expression in Escherichia coli. J Biotechnol. 115(2), 113-128, (2005).
3. Contreras-Gomez, A., Sanchez-Miron, A., Garcia-Camacho, F., Molina-Grima, E., & Chisti, Y. Protein production using the baculovirus-insect cell expression system. Biotechnol Prog. 30(1), 1-18, (2014).
4. Bandaranayake, A. D., & Almo, S. C. Recent advances in mammalian protein production. FEBS Lett. 588(2), 253-260, (2014).
5. Varki, A. Essentials of Glycobiology. Second edition edn, (Cold Spring Harbor NY), (2009)
6. Moremen, K. W., Tiemeyer, M., & Nairn, A. V. Vertebrate protein glycosylation: diversity, synthesis and function. Nat Rev Mol Cell Biol. 13(7), 448-462, (2012).
7. Apweiler, R., Hermjakob, H., & Sharon, N. On the frequency of protein glycosylation, as deduced from analysis of the SWISS-PROT database. Biochim Biophys Acta. 1473(1), 4-8, (1999).
8. Payne, R. J., & Wong, C. H. Advances in chemical ligation strategies for the synthesis of glycopeptides and glycoproteins. Chemical Comm. 46(1), 21-43, (2010).
9. Wang, P. et al. Erythropoietin derived by chemical synthesis. Science. 342(6164), 1357-1360, (2013).
10. Yamamoto, N., Tanabe, Y., Okamoto, R., Dawson, P. E., & Kajihara, Y. Chemical synthesis of a glycoprotein having an intact human complex- type sialyloligosaccharide under the Boc and Fmoc synthetic strategies. J Am Chem Soc. 130(2), 501-510, (2008).
11. Huang, W., Giddens, J., Fan, S. Q., Toonstra, C., & Wang, L. X. Chemoenzymatic glycoengineering of intact IgG antibodies for gain of functions. J Am Chem Soc. 134(29), 12308-12318, (2012).
12. Schwarz, F. et al. A combined method for producing homogeneous glycoproteins with eukaryotic N-glycosylation. Nat Chem Biol. 6(4), 264-266, (2010).
13. Smith, E. L. et al. Chemoenzymatic Fc glycosylation via engineered aldehyde tags. Bioconj Chem. 25(4), 788-795, (2014).
14. Zou, G. et al. Chemoenzymatic synthesis and Fcgamma receptor binding of homogeneous glycoforms of antibody Fc domain. Presence of a bisecting sugar moiety enhances the affinity of Fc to FcgammaIIIa receptor. J Am Chem Soc. 133(46), 18975-18991, (2011).
15. Cox, K. M. et al. Glycan optimization of a human monoclonal antibody in the aquatic plant Lemna minor. Nat Biotech. 24(12), 1591-1597, (2006).
16. Jarvis, D. L. Baculovirus-insect cell expression systems. Methods Enzymol. 463191-222, (2009)
17. Li, H. et al. Optimization of humanized IgGs in glycoengineered Pichia pastoris. Nature Biotech. 24(2), 210-215, (2006).
18. Palmberger, D., Wilson, I. B., Berger, I., Grabherr, R., & Rendic, D. SweetBac: a new approach for the production of mammalianised glycoproteins in insect cells. PLoS One. 7(4), e34226, (2012).
19. Toth, A. M., Kuo, C. W., Khoo, K. H., & Jarvis, D. L. A new insect cell glycoengineering approach provides baculovirus-inducible glycogene expression and increases human-type glycosylation efficiency. J Biotech. 182-183 (19-29), (2014).
20. Yamane-Ohnuki, N. et al. Establishment of FUT8 knockout Chinese hamster ovary cells: an ideal host cell line for producing completely defucosylated antibodies with enhanced antibody-dependent cellular cytotoxicity. Biotechnol Bioeng. 87(5), 614-622, (2004).
21. Dalton, A. C., & Barton, W. A. Over-expression of secreted proteins from mammalian cell lines. Protein Sci. 23(5), 517-525, (2014).
22. Legendre, J. Y., & Szoka, F. C., Jr. Cyclic amphipathic peptide-DNA complexes mediate high-efficiency transfection of adherent mammalian cells. Proc Natl Acad Sci U S A. 90(3), 893-897, (1993).
23. Wurm, F. M. Production of recombinant protein therapeutics in cultivated mammalian cells. Nat Biotech. 22(11), 1393-1398, (2004).
24. Jordan, M., Schallhorn, A., & Wurm, F. M. Transfecting mammalian cells: optimization of critical parameters affecting calcium-phosphate precipitate formation. Nucleic Acids Res. 24(4), 596-601, (1996).
25. Kingston, R. E., Chen, C. A., & Okayama, H. Calcium phosphate transfection. Curr Protoc Immunol. Chapter 10Unit 10 13, (2001)
26. Boussif, O. et al. A versatile vector for gene and oligonucleotide transfer into cells in culture and in vivo: polyethylenimine. Proc Natl Acad Sci U S A. 92(16), 7297-7301, (1995).
27. Pagano, J. S., McCutchan, J. H., & Vaheri, A. Factors influencing the enhancement of the infectivity of poliovirus ribonucleic acid by diethylaminoethyl-dextran. J Virol. 1(5), 891-897 (1967).
28. Fraley, R., Subramani, S., Berg, P., & Papahadjopoulos, D. Introduction of liposome-encapsulated SV40 DNA into cells. J Biol Chem. 255(21), 10431-10435 (1980).
29. Schlaeger, E. J., & Christensen, K. Transient gene expression in mammalian cells grown in serum-free suspension culture. Cytotechnology. 30(1-3), 71-83, (1999).
30. Longo, P. A., Kavran, J. M., Kim, M. S., & Leahy, D. J. Transient mammalian cell transfection with polyethylenimine (PEI). Methods Enz. 529227-240, (2013)
31. Backliwal, G., Hildinger, M., Hasija, V., & Wurm, F. M. High-density transfection with HEK-293 cells allows doubling of transient titers and removes need for a priori DNA complex formation with PEI. Biotechnol Bioeng. 99(3), 721-727, (2008).
32. Backliwal, G. et al. Rational vector design and multi-pathway modulation of HEK 293E cells yield recombinant antibody titers exceeding 1 g/l by transient transfection under serum-free conditions. Nucleic Acids Res. 36(15), e96, (2008).
33. Vink, T., Oudshoorn-Dickmann, M., Roza, M., Reitsma, J. J., & de Jong, R. N. A simple, robust and highly efficient transient expression system for producing antibodies. Methods. 65(1), 5-10, (2014).
34. Unson, C. G. Expression of glucagon receptors in tetracycline-inducible HEK293S GnT1- stable cell lines: an approach toward purification of receptor protein for structural studies. Biopolymers. 90(3), 287-296, (2008).
35. Reeves, P. J., Callewaert, N., Contreras, R., & Khorana, H. G. Structure and function in rhodopsin: high-level expression of rhodopsin with restricted and homogeneous N-glycosylation by a tetracycline-inducible N-acetylglucosaminyltransferase I-negative HEK293S stable mammalian cell line. Proc Natl Acad Sci U S A. 99(21), 13419-13424, (2002).
36. Stanley, P., Narasimhan, S., Siminovitch, L., & Schachter, H. Chinese hamster ovary cells selected for resistance to the cytotoxicity of phytohemagglutinin are deficient in a UDP-N-acetylglucosamine--glycoprotein N-acetylglucosaminyltransferase activity. Proc Natl Acad Sci U S A. 72(9), 3323-3327 (1975).
37. Schachter, H. The joys of HexNAc. The synthesis and function of N- and O-glycan branches. Glycoconjugate J. 17(7-9), 465-483, (2000).
38. Elbein, A. D., Tropea, J. E., Mitchell, M., & Kaushal, G. P. Kifunensine, a potent inhibitor of the glycoprotein processing mannosidase I. J Biol Chem. 265(26), 15599-15605 (1990).
39. Rillahan, C. D. et al. Global metabolic inhibitors of sialyl- and fucosyltransferases remodel the glycome. Nat Chem Biol. 8(7), 661-668, (2012).
40. Okeley, N. M. et al. Development of orally active inhibitors of protein and cellular fucosylation. Proc Natl Acad Sci U S A. 110(14), 5404-5409, (2013).
41. Chang, V. T. et al. Glycoprotein structural genomics: solving the glycosylation problem. Structure. 15(3), 267-273, (2007).
42. Barb, A. W. et al. NMR characterization of immunoglobulin G Fc glycan motion on enzymatic sialylation. Biochemistry. 51(22), 4618-4626, (2012).
43. Subedi, G. P., Hanson, Q. M., & Barb, A. W. Restricted motion of the conserved immunoglobulin G1 N-glycan is essential for efficient FcgammaRIIIa binding. Structure. 22(10), 1478-1488, (2014).
44. Barb, A. W., Brady, E. K., & Prestegard, J. H. Branch-specific sialylation of IgG-Fc glycans by ST6Gal-I. Biochemistry. 48(41), 9705-9707, (2009).
45. Anumula, K. R., & Taylor, P. B. A Comprehensive Procedure for Preparation of Partially Methylated Alditol Acetates from Glycoprotein Carbohydrates. Anal Biochem. 203(1), 101-108 (1992).
46. Parekh, R. B. et al. Association of rheumatoid arthritis and primary osteoarthritis with changes in the glycosylation pattern of total serum IgG. Nature. 316(6027), 452-457, (1985).
47. Yagi, H. et al. Backbone H, C, and N resonance assignments of the Fc fragment of human immunoglobulin G glycoprotein. Biomol NMR Assign., (2014).
48. Al-Rubeai, A. B. a. M. Effects of Serum and Growth Factor on HEK 293 Proliferation and Adenovirus Productivity. Animal Cell Tech: Basic & Appl Aspects. 15(19-23) (2009).
49. Kaufman, S. M. A. a. R. J. in New Comprehensive Biochemistry. Vol. 38 Gene Transfer and Expression in Mammalian Cells. (ed S. C. Makrides) Ch. 13, 411-432 Elsevier, (2003).
50. Berlec, A., & Strukelj, B. Current state and recent advances in biopharmaceutical production in Escherichia coli, yeasts and mammalian cells. J Ind Microbiol Biotechnol. 40(3-4), 257-274, (2013).
51. Nothaft, H., & Szymanski, C. M. Bacterial protein N-glycosylation: new perspectives and applications. J Biol Chem. 288(10), 6912-6920, (2013).
52. Barb, A. W. Intramolecular N-glycan/polypeptide interactions observed at multiple N-glycan remodeling steps through [(13)C,(15)N]-N- acetylglucosamine labeling of immunoglobulin G1. Biochemistry. 54(2), 313-322, (2015)