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Plant-Derived Biologics and Other High-Value Compounds
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Plant-Derived Biologics and Other High-Value Compounds

A special issue of International Journal of Molecular Sciences (ISSN 1422-0067). This special issue belongs to the section "Molecular Plant Sciences".

Deadline for manuscript submissions: closed (30 November 2022) | Viewed by 31015

Special Issue Editors

Prof. Dr. Ivan Minkov

E-Mail Website
Guest Editor
Institute of Molecular Biology and Biotechnologies (IMBB), 4000 Plovdiv, Bulgaria
Interests: systems biology; bioinformatics; plant and medical biotechnology; bionanotechnology, molecular farming
Dr. Gergana Zahmanova

E-Mail Website
Guest Editor
1. Center of Plant Systems Biology and Biotechnology, 4000 Plovdiv, Bulgaria
2. Department of Plant Physiology and Molecular Biology, University of Plovdiv, 4000 Plovdiv, Bulgaria
Interests: emerging viruses; vaccine design; antivirals; plant molecular farming; recombinant proteins; plant biotechnology
Special Issues, Collections and Topics in MDPI journals

Special Issue Information

Dear Colleagues,

Over the last twenty years, remarkable progress has been made in the field of plant-produced biologics. Soon we may even witness the first licensed plant-derived vaccines against influenza and SARS-CoV-2. Plant expression systems have a unique place among expression platforms more broadly. They allow plants to produce high-value recombinant biological compounds with high productivity and scalability, at а lower cost. Shortened R&D time, improved plant expression vectors, enhanced downstream processing, modified plant-glycosylation patterns to better match those observed in animals and humans – these aspects and more have helped advance the development of plant molecular farming (PMF) significantly.

This Special Issue of “Molecular Science” aims to highlight recent developments in and applications of plant expression systems and plant-derived biologics, as well as to evaluate their technical feasibility and economic implications.

Prof. Dr. Ivan Minkov
Dr. Gergana Zahmanova
Guest Editors

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Keywords

  • plant molecular farming (PMF)
  • biologics
  • vaccine candidates
  • monoclonal antibodies (mAbs)
  • diagnostic tools
  • virus-like particles (VLPs)
  • plant expression vectors
  • glycoengineering
  • posttranslational modification
  • economic impact evaluation

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Published Papers (6 papers)

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Research

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15 pages, 751 KiB  
Article
Sambucus ebulus (Elderberry) Fruits Modulate Inflammation and Complement System Activity in Humans
by Yoana Kiselova-Kaneva, Milka Nashar, Bogdan Roussev, Ayshe Salim, Minka Hristova, Pawel Olczyk, Katarzyna Komosinska-Vassev, Ivayla Dincheva, Ilian Badjakov, Bistra Galunska and Diana Ivanova
Int. J. Mol. Sci. 2023, 24(10), 8714; https://doi.org/10.3390/ijms24108714 - 13 May 2023
Cited by 3 | Viewed by 3022
Abstract
Sambucus ebulus (SE) fruits are used for immune stimulation and amelioration of gastrointestinal inflammatory conditions. Currently, there is no scientific evidence of their effects on various aspects of the immune response mechanisms in humans. The purpose of this study was to evaluate the [...] Read more.
Sambucus ebulus (SE) fruits are used for immune stimulation and amelioration of gastrointestinal inflammatory conditions. Currently, there is no scientific evidence of their effects on various aspects of the immune response mechanisms in humans. The purpose of this study was to evaluate the immunomodulatory potential of SE fruit infusion intake in healthy humans. Anthocyanin content was determined with UPLC-ESI-MS/MS. Fifty-three volunteers enrolled in a 4-week SE infusion intake intervention. Blood count, serum total protein, Interleukin 1 beta (IL-1β), Interleukin 6 (IL-6), Tumor Necrosis Factor Alpha (TNFα), High-sensitivity C-reactive protein (hs-CRP), C3, and C4 levels were measured on automatic analyzers, and Interleukin 8 (IL-8) was measured manually with an ELISA kit. Cyanidin-3-O-galactoside (48.15 mg/g DW), followed by cyaniding-3-sambubioside (43.41 ± 1.07 mg/g DW), were the most abundant anthocyanins in SE samples. A significant decrease in total protein (2.82%), IL-6 (20.15%), TNFα (5.38%), IL-8 (5.50%), C3 (4.16%), and C4 (14.29%) was established in the whole group. Total protein, IL-8, TNFα, and C4 decreased in women (3.11%, 4.76%, 5.09%, and 11.11%), and IL-6 decreased (40.61%) in men. Hb (1.20%) and hematocrit (1.55%) levels decreased in the whole group and in the women group (1.61% and 2.20%). SE fruits exert immune-modulatory activity as revealed by decreased pro-inflammatory status and complement activity markers in healthy volunteers after a 4-week intervention. Full article
(This article belongs to the Special Issue Plant-Derived Biologics and Other High-Value Compounds)
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16 pages, 3340 KiB  
Article
High-Yield Production of Chimeric Hepatitis E Virus-Like Particles Bearing the M2e Influenza Epitope and Receptor Binding Domain of SARS-CoV-2 in Plants Using Viral Vectors
by Eugenia S. Mardanova, Roman Y. Kotlyarov, Maya D. Stuchinskaya, Lyudmila I. Nikolaeva, Gergana Zahmanova and Nikolai V. Ravin
Int. J. Mol. Sci. 2022, 23(24), 15684; https://doi.org/10.3390/ijms232415684 - 10 Dec 2022
Cited by 11 | Viewed by 2934
Abstract
Capsid protein of Hepatitis E virus (HEV) is capable of self-assembly into virus-like particles (VLPs) when expressed in Nicotiana benthamiana plants. Such VLPs could be used as carriers of antigens for vaccine development. In this study, we obtained VLPs based on truncated coat [...] Read more.
Capsid protein of Hepatitis E virus (HEV) is capable of self-assembly into virus-like particles (VLPs) when expressed in Nicotiana benthamiana plants. Such VLPs could be used as carriers of antigens for vaccine development. In this study, we obtained VLPs based on truncated coat protein of HEV bearing the M2e peptide of Influenza A virus or receptor-binding domain of SARS-CoV-2 spike glycoprotein (RBD). We optimized the immunogenic epitopes’ presentation by inserting them into the protruding domain of HEV ORF2 at position Tyr485. The fusion proteins were expressed in Nicotiana benthamiana plants using self-replicating potato virus X (PVX)-based vector. The fusion protein HEV/M2, targeted to the cytosol, was expressed at the level of about 300–400 μg per gram of fresh leaf tissue and appeared to be soluble. The fusion protein was purified using metal affinity chromatography under native conditions with the final yield about 200 μg per gram of fresh leaf tissue. The fusion protein HEV/RBD, targeted to the endoplasmic reticulum, was expressed at about 80–100 μg per gram of fresh leaf tissue; the yield after purification was up to 20 μg per gram of fresh leaf tissue. The recombinant proteins HEV/M2 and HEV/RBD formed nanosized virus-like particles that could be recognized by antibodies against inserted epitopes. The ELISA assay showed that antibodies of COVID-19 patients can bind plant-produced HEV/RBD virus-like particles. This study shows that HEV capsid protein is a promising carrier for presentation of foreign antigen. Full article
(This article belongs to the Special Issue Plant-Derived Biologics and Other High-Value Compounds)
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18 pages, 2758 KiB  
Article
In Vitro Strategy for the Enhancement of the Production of Bioactive Polyphenols in Transformed Roots of Salvia bulleyana
by Marta Krzemińska, Aleksandra Owczarek, Monika A. Olszewska and Izabela Grzegorczyk-Karolak
Int. J. Mol. Sci. 2022, 23(14), 7771; https://doi.org/10.3390/ijms23147771 - 14 Jul 2022
Cited by 10 | Viewed by 1976
Abstract
The underground parts of Salvia bulleyana, a rare Chinese plant species, have long been used in traditional Chinese medicine. The Rhizobium rhizogenes-transformed root culture obtained from this plant might be a promising novel source of valuable phenolics, including rosmarinic acid. The [...] Read more.
The underground parts of Salvia bulleyana, a rare Chinese plant species, have long been used in traditional Chinese medicine. The Rhizobium rhizogenes-transformed root culture obtained from this plant might be a promising novel source of valuable phenolics, including rosmarinic acid. The present study identifies for the first time, the optimal growth conditions of S. bulleyana hairy roots regarding production efficiency. The comprehensive optimization comprised cultivation in different basal media (B5, SH, MS, and WP) with full- and half-strength macro- and microelements, different vitamin contents (full, half, one-quarter part, and without) and sucrose concentrations (2, 3, 4, 5%), and under different light conditions: in dark, under blue LED (λ = 430 nm), red LED (λ = 670 nm), mixed blue and red LED (30%:70%), and white LED (390–670 nm). Hairy root growth and bioactive compound accumulation were also detailed every five days over the 50-day culture cycle. The optimal conditions were determined using a technique for order preference by similarity to the ideal solution (TOPSIS). The most efficient combination for root growth and polyphenol content was found to be ½SH liquid medium with half vitamin concentration and 3% sucrose when grown in the dark. The biomass yield during the growth cycle was 6.1 g (fresh weight—FW) and 0.92 g (dry weight—DW) on one Erlenmeyer flask: a 14.3-fold increase in FW and 16.1-fold increase in DW in relation to the inoculum. The highest mean total phenolic content was 93.6 mg/g DW including about 70 mg/g DW rosmarinic acid, reached on day 40 of culture; compared to roots of two-year-old plants grown under field conditions, the total phenolic acid content was four times higher and rosmarinic acid eight times higher. The obtained results place the investigated culture among the best hair root cultures for rosmarinic acid production. Full article
(This article belongs to the Special Issue Plant-Derived Biologics and Other High-Value Compounds)
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Review

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29 pages, 1633 KiB  
Review
Green Biologics: Harnessing the Power of Plants to Produce Pharmaceuticals
by Gergana Zahmanova, Alaa A. A. Aljabali, Katerina Takova, George Minkov, Murtaza M. Tambuwala, Ivan Minkov and George P. Lomonossoff
Int. J. Mol. Sci. 2023, 24(24), 17575; https://doi.org/10.3390/ijms242417575 - 17 Dec 2023
Cited by 8 | Viewed by 6683
Abstract
Plants are increasingly used for the production of high-quality biological molecules for use as pharmaceuticals and biomaterials in industry. Plants have proved that they can produce life-saving therapeutic proteins (Elelyso™—Gaucher’s disease treatment, ZMapp™—anti-Ebola monoclonal antibodies, seasonal flu vaccine, Covifenz™—SARS-CoV-2 virus-like particle vaccine); however, [...] Read more.
Plants are increasingly used for the production of high-quality biological molecules for use as pharmaceuticals and biomaterials in industry. Plants have proved that they can produce life-saving therapeutic proteins (Elelyso™—Gaucher’s disease treatment, ZMapp™—anti-Ebola monoclonal antibodies, seasonal flu vaccine, Covifenz™—SARS-CoV-2 virus-like particle vaccine); however, some of these therapeutic proteins are difficult to bring to market, which leads to serious difficulties for the manufacturing companies. The closure of one of the leading companies in the sector (the Canadian biotech company Medicago Inc., producer of Covifenz) as a result of the withdrawal of investments from the parent company has led to the serious question: What is hindering the exploitation of plant-made biologics to improve health outcomes? Exploring the vast potential of plants as biological factories, this review provides an updated perspective on plant-derived biologics (PDB). A key focus is placed on the advancements in plant-based expression systems and highlighting cutting-edge technologies that streamline the production of complex protein-based biologics. The versatility of plant-derived biologics across diverse fields, such as human and animal health, industry, and agriculture, is emphasized. This review also meticulously examines regulatory considerations specific to plant-derived biologics, shedding light on the disparities faced compared to biologics produced in other systems. Full article
(This article belongs to the Special Issue Plant-Derived Biologics and Other High-Value Compounds)
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28 pages, 3992 KiB  
Review
The Plant Viruses and Molecular Farming: How Beneficial They Might Be for Human and Animal Health?
by Gergana Zahmanova, Alaa A. Aljabali, Katerina Takova, Valentina Toneva, Murtaza M. Tambuwala, Anton P. Andonov, Georgi L. Lukov and Ivan Minkov
Int. J. Mol. Sci. 2023, 24(2), 1533; https://doi.org/10.3390/ijms24021533 - 12 Jan 2023
Cited by 13 | Viewed by 9234
Abstract
Plant viruses have traditionally been studied as pathogens in the context of understanding the molecular and cellular mechanisms of a particular disease affecting crops. In recent years, viruses have emerged as a new alternative for producing biological nanomaterials and chimeric vaccines. Plant viruses [...] Read more.
Plant viruses have traditionally been studied as pathogens in the context of understanding the molecular and cellular mechanisms of a particular disease affecting crops. In recent years, viruses have emerged as a new alternative for producing biological nanomaterials and chimeric vaccines. Plant viruses were also used to generate highly efficient expression vectors, revolutionizing plant molecular farming (PMF). Several biological products, including recombinant vaccines, monoclonal antibodies, diagnostic reagents, and other pharmaceutical products produced in plants, have passed their clinical trials and are in their market implementation stage. PMF offers opportunities for fast, adaptive, and low-cost technology to meet ever-growing and critical global health needs. In this review, we summarized the advancements in the virus-like particles-based (VLPs-based) nanotechnologies and the role they played in the production of advanced vaccines, drugs, diagnostic bio-nanomaterials, and other bioactive cargos. We also highlighted various applications and advantages plant-produced vaccines have and their relevance for treating human and animal illnesses. Furthermore, we summarized the plant-based biologics that have passed through clinical trials, the unique challenges they faced, and the challenges they will face to qualify, become available, and succeed on the market. Full article
(This article belongs to the Special Issue Plant-Derived Biologics and Other High-Value Compounds)
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17 pages, 1206 KiB  
Review
Maximizing the Production of Recombinant Proteins in Plants: From Transcription to Protein Stability
by Ziru Feng, Xifeng Li, Baofang Fan, Cheng Zhu and Zhixiang Chen
Int. J. Mol. Sci. 2022, 23(21), 13516; https://doi.org/10.3390/ijms232113516 - 4 Nov 2022
Cited by 19 | Viewed by 5638
Abstract
The production of therapeutic and industrial recombinant proteins in plants has advantages over established bacterial and mammalian systems in terms of cost, scalability, growth conditions, and product safety. In order to compete with these conventional expression systems, however, plant expression platforms must have [...] Read more.
The production of therapeutic and industrial recombinant proteins in plants has advantages over established bacterial and mammalian systems in terms of cost, scalability, growth conditions, and product safety. In order to compete with these conventional expression systems, however, plant expression platforms must have additional economic advantages by demonstrating a high protein production yield with consistent quality. Over the past decades, important progress has been made in developing strategies to increase the yield of recombinant proteins in plants by enhancing their expression and reducing their degradation. Unlike bacterial and animal systems, plant expression systems can utilize not only cell cultures but also whole plants for the production of recombinant proteins. The development of viral vectors and chloroplast transformation has opened new strategies to drastically increase the yield of recombinant proteins from plants. The identification of promoters for strong, constitutive, and inducible promoters or the tissue-specific expression of transgenes allows for the production of recombinant proteins at high levels and for special purposes. Advances in the understanding of RNAi have led to effective strategies for reducing gene silencing and increasing recombinant protein production. An increased understanding of protein translation, quality control, trafficking, and degradation has also helped with the development of approaches to enhance the synthesis and stability of recombinant proteins in plants. In this review, we discuss the progress in understanding the processes that control the synthesis and degradation of gene transcripts and proteins, which underlie a variety of developed strategies aimed at maximizing recombinant protein production in plants. Full article
(This article belongs to the Special Issue Plant-Derived Biologics and Other High-Value Compounds)
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