Easily purified baculovirus/insect-system-expressed recombinant hepatitis B virus surface antigen fused to the N- or C-terminus of polyhedrin

Silva, Leonardo A.; Camargo, Brenda Rabelo de; Araújo, Ana Carolina; Batista, Taylice Leonel; Ribeiro, Bergmann Morais; Ardisson-Araújo, Daniel Mendes Pereira

doi:10.1007/s00705-021-05305-6

Cited by 3 publications

(2 citation statements)

References 36 publications

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“…*P < 0.05 and **P < 0.01, compared with the control group. (Fernandes et al 2022;Silva et al 2022). To improve the CD200-Fc protein expression yield, an endoplasmic reticulum retention peptide (KDEL) was tagged at the CD200-Fc Cterminus (Newstead & Barr 2020).…”

Section: Discussionmentioning

confidence: 99%

Insect cell‐derived human CD200‐Fc increases zonula occludens‐1 tight junction protein in urothelial carcinoma cells

Lee,

Lee

et al. 2023

Entomological Research

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CD200 is a ligand that interacts with the CD200 receptor 1, with an anti‐inflammatory effect. CD200 inhibits the nuclear factor kappa‐light‐chain‐enhancer of activated B cells (NF‐κB) pathway, attenuating the inflammatory response and maintaining barrier function. In this study, human CD200 was fused with the fragment crystallizable (Fc) region of human immunoglobulin G (IgG) to generate the recombinant CD200‐Fc protein, produced in insect cells using the baculovirus expression vector system. The CD200‐Fc gene was cloned under the control of the polyhedrin promoter in the pFastBac‐1 vector of the baculovirus expression vector system. Immunoblot analysis confirmed the expression of CD200‐Fc in insect cells. The CD200‐Fc protein was successfully purified using protein A affinity chromatography. 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide (MTT) assay revealed that purified CD200‐Fc protein inhibited the proliferation of TCCSUP cells, a bladder epithelial cancer cell expressing CD200 receptor 1, at concentrations of 1, 10 and 100 ng/mL. Quantitative real‐time reverse transcription PCR (qRT‐PCR) and immunoblot analyses confirmed that the mRNA and protein levels of zonula occludens‐1, a tight junction protein for barrier protection in epithelial tissues, were increased in TCCSUP cells treated with insect cell‐derived CD200‐Fc. These results suggest that insect cell‐derived CD200‐Fc could alter zonula occludens‐1 expression in bladder epithelial cancer cells, enhancing the function of the cell barrier protein, which could inhibit cancer metastasis. Taken together, the baculovirus expression vector system can be applied to express antitumor therapeutic CD200‐Fc protein for the inhibition of bladder cancer.

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Section: Discussionmentioning

confidence: 99%

Insect cell‐derived human CD200‐Fc increases zonula occludens‐1 tight junction protein in urothelial carcinoma cells

Lee,

Lee

et al. 2023

Entomological Research

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“…The CHIKV multi-epitope gene (R1) that contains an 18 amino acid sequence from the E2 region and a sequence of 17 amino acids from the nsP3 region ( Figure 1 A) was synthetized by the company, Integrated DNA Technologies (IDT). The R1 gene was cloned into the Nco I restriction enzyme site of the vector pFastBac1-6xhis-polh [ 36 , 37 ]. After that, the donor pFastBac1-6xhis-R1-polh was transformed in E. coli DH10Bac cells (Thermo Fisher Scientific, Waltham, MA, USA) by electroporation and the R1 gene, under control of the polh promoter, was inserted into the genome of Autographa californica multiple nucleopolyhedrovirus (AcMNPV) via transposition (Bac-to-Bac ® , Baculovirus Expression Systems, Thermo Fisher Scientific, Waltham, MA, USA), generating the recombinant bacmid vAc-R1-6xHis.…”

Section: Methodsmentioning

confidence: 99%

A Chikungunya Virus Multiepitope Recombinant Protein Expressed from the Binary System Insect Cell/Recombinant Baculovirus Is Useful for Laboratorial Diagnosis of Chikungunya

et al. 2022

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Chikungunya virus (CHIKV) is an arbovirus currently distributed worldwide, causing a disease that shares clinical signs and symptoms with other illnesses, such as dengue and Zika and leading to a challenging clinical differential diagnosis. In Brazil, CHIKV emerged in 2014 with the simultaneous introduction of both Asian and East/Central/South African (ECSA) genotypes. Laboratorial diagnosis of CHIKV is mainly performed by molecular and serological assays, with the latter more widely used. Although many commercial kits are available, their costs are still high for many underdeveloped and developing countries where the virus circulates. Here we described the development and evaluation of a multi-epitope recombinant protein-based IgG-ELISA (MULTREC IgG-ELISA) test for the specific detection of anti-CHIKV antibodies in clinical samples, as an alternative approach for laboratorial diagnosis. The MULTREC IgG-ELISA showed 86.36% of sensitivity and 100% of specificity, and no cross-reactivity with other exanthematic diseases was observed. The recombinant protein was expressed from the binary system insect cell/baculovirus using the crystal-forming baculoviral protein polyhedrin as a carrier of the target recombinant protein to facilitate recovery. The crystals were at least 10 times smaller in size and had an amorphous shape when compared to the polyhedrin wild-type crystal. The assay uses a multi-epitope antigen, representing two replicates of 18 amino acid sequences from the E2 region and a sequence of 17 amino acids from the nsP3 region of CHIKV. The recombinant protein was highly expressed, easy to purify and has demonstrated its usefulness in confirming chikungunya exposure, indeed showing a good potential tool for epidemiological surveillance.

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Comparison between different conditions for the incorporation of foreign proteins into Autographa californica multiple nucleopolyhedrovirus polyhedra for biotechnological purposes

López,

Gravisaco

et al. 2024

Arch Virol

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Easily purified baculovirus/insect-system-expressed recombinant hepatitis B virus surface antigen fused to the N- or C-terminus of polyhedrin

Cited by 3 publications

References 36 publications

Insect cell‐derived human CD200‐Fc increases zonula occludens‐1 tight junction protein in urothelial carcinoma cells

Insect cell‐derived human CD200‐Fc increases zonula occludens‐1 tight junction protein in urothelial carcinoma cells

A Chikungunya Virus Multiepitope Recombinant Protein Expressed from the Binary System Insect Cell/Recombinant Baculovirus Is Useful for Laboratorial Diagnosis of Chikungunya

Comparison between different conditions for the incorporation of foreign proteins into Autographa californica multiple nucleopolyhedrovirus polyhedra for biotechnological purposes

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