Protein Expression and Purification, volume 183, pages 105864

A simple method to purify recombinant HCV core protein expressed in Pichia pastoris for obtaining virus-like particles and producing monoclonal antibodies.

Pechelyulko Anastasia ¹

Sokolova Olga S.

Andreeva Kovalevskaya Zhanna ²

Dmitriev Dmitriy

Lavrov Viacheslav ¹

Massino Yulia ¹

Nagel Alexey ²

Segal Olga ¹

Solonin Alexander

Tarakanova Yulia ¹

Dmitriev Alexander

Dmitriev D ¹

Sokolova Olga S. ³

Solonin A. ²

Dmitriev Alexander K. ¹

Hide authors affiliations Show authors affiliations: 3 affiliations

Mechnikov Scientific Research Institute of Vaccines and Sera, 5A Maly Kazenny Lane, Moscow, 105064, Russia. |

FSBIS G. K. Skryabin Institute of Biochemistry and Physiology of Microorganisms, Russian Academy of Sciences, Pushchino, Moscow Region, Russia |

Department of Bioengineering, Faculty of Biology, Lomonosov Moscow State University, Moscow, 119991, Russia |

Publication type: Journal Article

Publication date: 2021-07-01

Elsevier

Journal: Protein Expression and Purification

Quartile SCImago

Quartile WOS

Impact factor: 1.6

ISSN: 10465928, 10960279

DOI: 10.1016/j.pep.2021.105864

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Biotechnology

Abstract

In this study, we describe an optimized method of obtaining virus-like particles (VLPs) of the recombinant hepatitis C virus (HCV) core protein (HCcAg) expressed in yeast cells (Pichia pastoris), which can be used for the construction of diagnostic test systems and vaccine engineering. The described simplified procedure was developed to enable in vitro self-assembly of HCcAg molecules into VLPs during protein purification. In brief, the HCcAg protein was precipitated from yeast cell lysates with ammonium sulfate and renatured by gel filtration on Sephadex G-25 under reducing conditions. VLPs were self-assembled after the removal of the reducing agent by gel filtration on Sephadex G-25. Protein purity and specificity were evaluated by SDS-PAGE and immunoblotting analysis. The molecular mass of VLPs and their relative quantity were measured by HPLC, followed by confirmation of VLPs production and estimation of their shape and size by transmission electron microscopy. As a result, we obtained recombinant HCcAg preparation (with ~90% purity) in the form of VLPs and monomers, which has been used to produce hybridomas secreting monoclonal antibodies (mAbs) against HCcAg.

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Vaccines	Vaccines, 1, 25% Vaccines 1 publication, 25%
Experimental and Therapeutic Medicine	Experimental and Therapeutic Medicine, 1, 25% Experimental and Therapeutic Medicine 1 publication, 25%
Applied Microbiology	Applied Microbiology, 1, 25% Applied Microbiology 1 publication, 25%
Therapeutic Delivery	Therapeutic Delivery, 1, 25% Therapeutic Delivery 1 publication, 25%
	1

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Multidisciplinary Digital Publishing Institute (MDPI)	Multidisciplinary Digital Publishing Institute (MDPI), 1, 25% Multidisciplinary Digital Publishing Institute (MDPI) 1 publication, 25%
Spandidos Publications	Spandidos Publications, 1, 25% Spandidos Publications 1 publication, 25%
American Society for Microbiology	American Society for Microbiology, 1, 25% American Society for Microbiology 1 publication, 25%
Future Medicine	Future Medicine, 1, 25% Future Medicine 1 publication, 25%
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Pechelyulko A. et al. A simple method to purify recombinant HCV core protein expressed in Pichia pastoris for obtaining virus-like particles and producing monoclonal antibodies. // Protein Expression and Purification. 2021. Vol. 183. p. 105864.

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Pechelyulko A., Andreeva Kovalevskaya Z., Dmitriev D., Lavrov V., Massino Y., Nagel A., Segal O., Sokolova O. S., Solonin A., Tarakanova Y., Dmitriev A., Dmitriev D., Sokolova O. S., Solonin A., Dmitriev A. K. A simple method to purify recombinant HCV core protein expressed in Pichia pastoris for obtaining virus-like particles and producing monoclonal antibodies. // Protein Expression and Purification. 2021. Vol. 183. p. 105864.

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RIS Copy

TY - JOUR

DO - 10.1016/j.pep.2021.105864

UR - https://doi.org/10.1016%2Fj.pep.2021.105864

TI - A simple method to purify recombinant HCV core protein expressed in Pichia pastoris for obtaining virus-like particles and producing monoclonal antibodies.

T2 - Protein Expression and Purification

AU - Pechelyulko, Anastasia

AU - Andreeva Kovalevskaya, Zhanna

AU - Dmitriev, Dmitriy

AU - Lavrov, Viacheslav

AU - Massino, Yulia

AU - Nagel, Alexey

AU - Segal, Olga

AU - Sokolova, Olga S.

AU - Solonin, Alexander

AU - Tarakanova, Yulia

AU - Dmitriev, Alexander

AU - Dmitriev, D

AU - Sokolova, Olga S.

AU - Solonin, A.

AU - Dmitriev, Alexander K.

PY - 2021

DA - 2021/07/01 00:00:00

PB - Elsevier

SP - 105864

VL - 183

SN - 1046-5928

SN - 1096-0279

ER -

BibTex

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BibTex Copy

@article{2021_Pechelyulko,

author = {Anastasia Pechelyulko and Zhanna Andreeva Kovalevskaya and Dmitriy Dmitriev and Viacheslav Lavrov and Yulia Massino and Alexey Nagel and Olga Segal and Olga S. Sokolova and Alexander Solonin and Yulia Tarakanova and Alexander Dmitriev and D Dmitriev and Olga S. Sokolova and A. Solonin and Alexander K. Dmitriev},

title = {A simple method to purify recombinant HCV core protein expressed in Pichia pastoris for obtaining virus-like particles and producing monoclonal antibodies.},

journal = {Protein Expression and Purification},

year = {2021},

volume = {183},

publisher = {Elsevier},

month = {jul},

url = {https://doi.org/10.1016%2Fj.pep.2021.105864},

pages = {105864},

doi = {10.1016/j.pep.2021.105864}

}

Found error?

Publisher

Elsevier

Journal

Protein Expression and Purification

Quartile SCImago

Quartile WOS

Impact factor

1.6

ISSN

10465928 (Print)
10960279 (Electronic)

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