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      Co-expression vs. co-infection using baculovirus expression vectors in insect cell culture: Benefits and drawbacks

      review-article
      1 , 1 , , , , * , 1
      Biotechnology Advances
      Elsevier Inc.
      AAV, adeno-associated virus, AcMNPV, Autographa californica multiple nucleopolyhedrovirus, ATP, adenosine triphosphate, BEVS, baculovirus expression vector system, BiP, binding immunoglobin protein, Cap, capsid, CLP, core-like particle, CMP-SAS, CMP-sialic acid synthase, CYP, cytochrome P, DNA, deoxyribonucleic acid, ER, endoplasmic reticulum, ETL, early-to-late, FISH, fluorescence in-situ hybridization, GFP, green fluorescent protein, HIV, human immunodeficiency virus, HR, homologous regions, HPV, human papillomavirus, MAP, mitogen-activated protein, MKK1, MAP kinase kinase 1 (MKK1 or MEK1), MAP3K, MAP kinase kinase kinase (MAP3K or MEKK), MOI, multiplicity of infection, mRNA, messenger RNA, MRP, multidrug resistance protein, PDI, protein disulfide isomerase, P-gp, P-glycoprotein, Polh, polyhedrin, OR, oxidoreductase, SAS, sialic acid phosphate synthase, SIV, simian immunodeficiency virus, PCR, polymerase chain reaction, Rep, replication, RNA, ribonucleic acid, rRNA, ribosomal RNA, STAT, signal transducers and activators of transcription, SUMO, small ubiquitin-like modifier, TOI, time of infection, ΔTOI, staggered time of infection, tPA, tissue plasminogen activator, VLP, virus-like particle, VP, virus protein, Baculovirus, Co-infection, Co-expression, Virus-like particle, Cytochrome P450, Chaperone, Promoter, Vector, Insect cell, Heterologous protein

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          Abstract

          The baculovirus expression vector system (BEVS) is a versatile and powerful platform for protein expression in insect cells. With the ability to approach similar post-translational modifications as in mammalian cells, the BEVS offers a number of advantages including high levels of expression as well as an inherent safety during manufacture and of the final product. Many BEVS products include proteins and protein complexes that require expression from more than one gene. This review examines the expression strategies that have been used to this end and focuses on the distinguishing features between those that make use of single polycistronic baculovirus (co-expression) and those that use multiple monocistronic baculoviruses (co-infection). Three major areas in which researchers have been able to take advantage of co-expression/co-infection are addressed, including compound structure-function studies, insect cell functionality augmentation, and VLP production. The core of the review discusses the parameters of interest for co-infection and co-expression with time of infection (TOI) and multiplicity of infection (MOI) highlighted for the former and the choice of promoter for the latter. In addition, an overview of modeling approaches is presented, with a suggested trajectory for future exploration. The review concludes with an examination of the gaps that still remain in co-expression/co-infection knowledge and practice.

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          Most cited references207

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          Molecular chaperones in cellular protein folding.

          F U Hartl (1996)
          The folding of many newly synthesized proteins in the cell depends on a set of conserved proteins known as molecular chaperones. These prevent the formation of misfolded protein structures, both under normal conditions and when cells are exposed to stresses such as high temperature. Significant progress has been made in the understanding of the ATP-dependent mechanisms used by the Hsp70 and chaperonin families of molecular chaperones, which can cooperate to assist in folding new polypeptide chains.
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            Real-time PCR for mRNA quantitation

            Real-time PCR has become one of the most widely used methods of gene quantitation because it has a large dynamic range, boasts tremendous sensitivity, can be highly sequence-specific, has little to no post-amplification processing, and is amenable to increasing sample throughput. However, optimal benefit from these advantages requires a clear understanding of the many options available for running a real-time PCR experiment. Starting with the theory behind real-time PCR, this review discusses the key components of a real-time PCR experiment, including one-step or two-step PCR, absolute versus relative quantitation, mathematical models available for relative quantitation and amplification efficiency calculations, types of normalization or data correction, and detection chemistries. In addition, the many causes of variation as well as methods to calculate intra- and inter-assay variation are addressed.
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              Baculovirus expression system for heterologous multiprotein complexes.

              The discovery of large multiprotein complexes in cells has increased the demand for improved heterologous protein production techniques to study their molecular structure and function. Here we describe MultiBac, a simple and versatile system for generating recombinant baculovirus DNA to express protein complexes comprising many subunits. Our method uses transfer vectors containing a multiplication module that can be nested to facilitate assembly of polycistronic expression cassettes, thereby minimizing requirements for unique restriction sites. The transfer vectors access a modified baculovirus DNA through Cre-loxP site-specific recombination or Tn7 transposition. This baculovirus has improved protein expression characteristics because specific viral genes have been eliminated. Gene insertion reactions are carried out in Escherichia coli either sequentially or concurrently in a rapid, one-step procedure. Our system is useful for both recombinant multiprotein production and multigene transfer applications.
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                Author and article information

                Contributors
                Journal
                Biotechnol Adv
                Biotechnol. Adv
                Biotechnology Advances
                Elsevier Inc.
                0734-9750
                1873-1899
                26 January 2012
                May-June 2012
                26 January 2012
                : 30
                : 3
                : 766-781
                Affiliations
                Department of Chemical Engineering, Waterloo Institute for Nanotechnology, University of Waterloo, Waterloo, Ontario, Canada N2L 3G1
                Author notes
                [* ]Corresponding author at: Department of Chemical Engineering, University of Waterloo, 200 University Ave. W, Waterloo, Ontario, Canada N2L 3G1. maucoin@ 123456uwaterloo.ca
                [1]

                Denotes equal contribution.

                Article
                S0734-9750(12)00011-0
                10.1016/j.biotechadv.2012.01.009
                7132753
                22297133
                4c306ade-a03f-4516-b3d9-040c185a763b
                Copyright © 2012 Elsevier Inc. All rights reserved.

                Since January 2020 Elsevier has created a COVID-19 resource centre with free information in English and Mandarin on the novel coronavirus COVID-19. The COVID-19 resource centre is hosted on Elsevier Connect, the company's public news and information website. Elsevier hereby grants permission to make all its COVID-19-related research that is available on the COVID-19 resource centre - including this research content - immediately available in PubMed Central and other publicly funded repositories, such as the WHO COVID database with rights for unrestricted research re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for free by Elsevier for as long as the COVID-19 resource centre remains active.

                History
                : 28 September 2011
                : 13 January 2012
                : 17 January 2012
                Categories
                Article

                Biotechnology
                aav, adeno-associated virus,acmnpv, autographa californica multiple nucleopolyhedrovirus,atp, adenosine triphosphate,bevs, baculovirus expression vector system,bip, binding immunoglobin protein,cap, capsid,clp, core-like particle,cmp-sas, cmp-sialic acid synthase,cyp, cytochrome p,dna, deoxyribonucleic acid,er, endoplasmic reticulum,etl, early-to-late,fish, fluorescence in-situ hybridization,gfp, green fluorescent protein,hiv, human immunodeficiency virus,hr, homologous regions,hpv, human papillomavirus,map, mitogen-activated protein,mkk1, map kinase kinase 1 (mkk1 or mek1),map3k, map kinase kinase kinase (map3k or mekk),moi, multiplicity of infection,mrna, messenger rna,mrp, multidrug resistance protein,pdi, protein disulfide isomerase,p-gp, p-glycoprotein,polh, polyhedrin,or, oxidoreductase,sas, sialic acid phosphate synthase,siv, simian immunodeficiency virus,pcr, polymerase chain reaction,rep, replication,rna, ribonucleic acid,rrna, ribosomal rna,stat, signal transducers and activators of transcription,sumo, small ubiquitin-like modifier,toi, time of infection,δtoi, staggered time of infection,tpa, tissue plasminogen activator,vlp, virus-like particle,vp, virus protein,baculovirus,co-infection,co-expression,virus-like particle,cytochrome p450,chaperone,promoter,vector,insect cell,heterologous protein

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