Multiplicity of infection and culture medium on the SARS-CoV-2 virus like-particles production by baculovirus/insect system.

IF 2 4区生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Biotechnology Letters Pub Date : 2025-03-06 DOI:10.1007/s10529-025-03572-w

Luis Giovani de Oliveira Guardalini, Felipe Moura Dias, Samanta Omae Camalhonte, Jaci Leme, Thaissa Consoni Bernardino, Felipe Soares Sposito, Eduardo Dias, Renato Mancini Astray, Aldo Tonso, Soraia Attie Calil Jorge, Eutimio Gustavo Fernández Núñez

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Abstract

This work aimed to assess the SARS-CoV-2 structural proteins' expression and virus-like particles (VLP) production by Baculovirus/Insect cell platform using two levels of Multiplicity of Infection (MOI), and two culture media, one of them a serum-free medium and the other one chemically defined. Two SARS-CoV-2 VLP were obtained from Sf9 cells coinfection using in both cases, three monocistronic recombinant baculoviruses holding the genes of Nucleocapsid (N; MOI = 2 or 0.2), Membrane (M; MOI = 1 or 0.1), and Envelope (E; MOI = 1 or 0.1) viral proteins, and the fourth one was changed between a baculovirus bearing Spike protein (S; MOI = 3 or 0.3) or receptor-binding domain (RBD; MOI = 3 or 0.3) genes of SARS-CoV-2. Similar performance was verified for both culture media in SARS-CoV-2 VLP production bearing four structural virus proteins or RBD domain. The SARS-CoV-2 structural proteins' expression was comparable at different MOIs (tenfold) as well as SARS-CoV-2 VLP size (around 100 nm). The increase in specific death rates over the coinfection phase was confirmed in relatively high MOI assays. This finding was related to an exponential virus titer profile for high MOIs over the entire infection phase, meanwhile, a viral peak was observed at low MOIs, confirming a secondary infection. The SARS-CoV-2 VLP improved production carrying immunogenic S protein was confirmed concerning others holding RBD. However, the protein composition of produced VLP should be studied further to assess the VLP homogeneity when different culture media and MOIs are used.

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杆状病毒/昆虫系统生产SARS-CoV-2病毒样颗粒的感染和培养基多样性

本工作旨在利用两种感染多重性（Multiplicity of Infection， MOI）水平和两种培养基（一种为无血清培养基，另一种为化学定义培养基），评估杆状病毒/昆虫细胞平台上SARS-CoV-2结构蛋白的表达和病毒样颗粒（VLP）的产生。从Sf9细胞共感染中获得了两个SARS-CoV-2 VLP，在这两种情况下，三种单顺性重组杆状病毒携带核衣壳(N；MOI = 2或0.2)，膜(M；MOI = 1或0.1),Envelope (E；MOI = 1或0.1)病毒蛋白，第四种是携带Spike蛋白的杆状病毒(S；MOI = 3或0.3)或受体结合域(RBD；MOI = 3或0.3)个SARS-CoV-2基因。两种培养基在含有四种结构病毒蛋白或RBD结构域的SARS-CoV-2 VLP生产中验证了相似的性能。SARS-CoV-2结构蛋白在不同MOIs（10倍）和SARS-CoV-2 VLP大小（约100 nm）下的表达具有可比性。在相对较高的MOI测定中证实了合并感染阶段特异性死亡率的增加。这一发现与整个感染阶段的高moi病毒滴度指数谱有关，同时，在低moi时观察到病毒峰值，证实了继发性感染。SARS-CoV-2 VLP提高了携带免疫原性S蛋白的产量，在其他RBD患者中得到证实。但是，在不同培养基和MOIs的情况下，需要进一步研究所得VLP的蛋白质组成，以评估VLP的均匀性。

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来源期刊

Biotechnology Letters 工程技术-生物工程与应用微生物

CiteScore

5.90

自引率

3.70%

发文量

108

审稿时长

1.2 months

期刊介绍： Biotechnology Letters is the world’s leading rapid-publication primary journal dedicated to biotechnology as a whole – that is to topics relating to actual or potential applications of biological reactions affected by microbial, plant or animal cells and biocatalysts derived from them. All relevant aspects of molecular biology, genetics and cell biochemistry, of process and reactor design, of pre- and post-treatment steps, and of manufacturing or service operations are therefore included. Contributions from industrial and academic laboratories are equally welcome. We also welcome contributions covering biotechnological aspects of regenerative medicine and biomaterials and also cancer biotechnology. Criteria for the acceptance of papers relate to our aim of publishing useful and informative results that will be of value to other workers in related fields. The emphasis is very much on novelty and immediacy in order to justify rapid publication of authors’ results. It should be noted, however, that we do not normally publish papers (but this is not absolute) that deal with unidentified consortia of microorganisms (e.g. as in activated sludge) as these results may not be easily reproducible in other laboratories. Papers describing the isolation and identification of microorganisms are not regarded as appropriate but such information can be appended as supporting information to a paper. Papers dealing with simple process development are usually considered to lack sufficient novelty or interest to warrant publication.