Paving the way for the production of secretory proteins by yeast cell factories

Huertas, María José; Michán, Carmen

doi:10.1111/1751-7915.13342

Cited by 14 publications

(5 citation statements)

References 11 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Yeasts are good choices as expression hosts for recombinant proteins due to their rapid growth, easy genetic manipulation, cost-effective growth medium requirements, available complete genome sequences, and ability to provide PTMs (Fletcher et al, 2016;Vieira Gomes et al, 2018;Baghban et al, 2019;Huertas and Michán, 2019). Codon bias and extracellular expressions occur with the recombinant proteins expressed using this system.…”

Section: Yeastmentioning

confidence: 99%

Recent Developments in Bioprocessing of Recombinant Proteins: Expression Hosts and Process Development

Tripathi

Shrivastava

2019

Front. Bioeng. Biotechnol.

369

243

View full text Add to dashboard Cite

Infectious diseases, along with cancers, are among the main causes of death among humans worldwide. The production of therapeutic proteins for treating diseases at large scale for millions of individuals is one of the essential needs of mankind. Recent progress in the area of recombinant DNA technologies has paved the way to producing recombinant proteins that can be used as therapeutics, vaccines, and diagnostic reagents. Recombinant proteins for these applications are mainly produced using prokaryotic and eukaryotic expression host systems such as mammalian cells, bacteria, yeast, insect cells, and transgenic plants at laboratory scale as well as in large-scale settings. The development of efficient bioprocessing strategies is crucial for industrial production of recombinant proteins of therapeutic and prophylactic importance. Recently, advances have been made in the various areas of bioprocessing and are being utilized to develop effective processes for producing recombinant proteins. These include the use of high-throughput devices for effective bioprocess optimization and of disposable systems, continuous upstream processing, continuous chromatography, integrated continuous bioprocessing, Quality by Design, and process analytical technologies to achieve quality product with higher yield. This review summarizes recent developments in the bioprocessing of recombinant proteins, including in various expression systems, bioprocess development, and the upstream and downstream processing of recombinant proteins.

show abstract

Section: Yeastmentioning

confidence: 99%

Recent Developments in Bioprocessing of Recombinant Proteins: Expression Hosts and Process Development

Tripathi

Shrivastava

2019

Front. Bioeng. Biotechnol.

369

243

View full text Add to dashboard Cite

show abstract

“…Yeasts are commonly utilized as host expression systems due to their fast growth rate, similar but less than bacteria, predisposition to genetic manipulations, existing genetic information, low-cost fermentation characteristics, and capability of providing proper post-translational modifications. Saccharomyces cerevisiae and Pichia pastoris are the most widely used yeast expression systems for biologics manufacturing (11)(12)(13). Various genes associated with secretion and Golgi trafficking of proteins have been modified in S.cerevisiae to increase the productivity up to a gram per liter level (14).…”

Section: General Introduction and Expression Systems Used In Bioproce...mentioning

confidence: 99%

Mammalian cell lines used in bioprocessing

ERGEN¹,

TÜFEKÇİ²

2022

J. Exp. Clin. Med.

View full text Add to dashboard Cite

A various number of expressions and host systems are used in biologics manufacturing. The most commonly preferred systems are based on bacteria, yeast, mammalian cells, insect cells, and transgenic animals. A wide range of molecules, including insulin, mAbs, vaccines, and recombinant proteins, are produced using different host systems. Because of several reasons impacting the product quality and yield, mammalian cells are utilized. Moreover, mammalian cells are generally used in virus-based vaccine manufacturing. Chinese Hamster Ovary (CHO) is the most widely used cell line for high yield stable recombinant protein production, while Human Embryonic Kidney (HEK) is favoured for transient transfection low yield protein manufacturing, viral-based vaccine and gene and cell therapy-related vector production. Other mammalian cell lines such as NSO, Sp2.0, Vero, MRC-5 and PerC.6 are also used in both recombinant protein and virus productions. Multiple modifications are carried out on industrial cell lines to make them more suitable for high yield and high-quality protein production. Thanks to these alterations, high productivity and quality levels are achieved in the biotechnology industry.

show abstract

“…Hence, the current scenario redirects to the development of alternative strategies for the production of such molecules in high yields using safe and economical technologies (Siddiqui et al, 2020). With the advent of genetic engineering, various platforms, such as those based on bacteria (Huang et al, 2012;Gupta and Shukla, 2016;Demain and Vaishnav, 2009), yeast (Ahmad et al, 2014;Kim et al, 2015;Baghban et al, 2019;Huertas and Michań, 2019), mammalian cells (Hacker and Balasubramanian, 2016;Gupta et al, 2017;Owczarek et al, 2019;O'Flaherty et al, 2020), insect cells (Contreras-Goḿez et al, 2014;Felberbaum, 2015;Kost and Kemp, 2016), plants (Shanmugaraj and Ramalingam, 2014;Lomonossoff and D'Aoust, 2016;Shanmugaraj et al, 2020), and transgenic animals (Moura et al, 2011;Maksimenko et al, 2013), have been largely and profoundly studied to explore the potential to develop them as mature platforms for the heterologous expression of proteins that have commercial and industrial importance (Merlin et al, 2014). However, although advances in the use of expression hosts for sustainable bioprocesses for the production of high-value products have been achieved, many challenges still need to be overcome in this field.…”

Section: Introductionmentioning

confidence: 99%

Construction and Validation of a Chloroplast Expression Vector for the Production of Recombinant Proteins in Chlorella vulgaris

Bolaños-Martínez¹,

Malla²,

Rosales‐Mendoza³

et al. 2022

Front. Mar. Sci.

View full text Add to dashboard Cite

Microalgae constitute a diverse group of photosynthetic unicellular microorganisms that have gained immense attention for biotechnological applications. They are promising platforms for the production of high-value metabolites and biopharmaceuticals for commercial and therapeutic applications because of their physiological properties and capability to grow easily in natural and artificial environments. Although the proof-of-concept for some applications have been achieved for model species, such as Chlamydomonas reinhardtii, the genetic engineering methods for microalgae are still in their infancy. Thus, an expansion of this field is required. Chlorella vulgaris is an important algal species with a high protein content and requires focus for the development of an efficient nuclear and chloroplast transformation process. This research aimed to establish a chloroplast transformation method for the freshwater green-algae species C. vulgaris based on a specific expression vector (pCMCC, which was named after Chula Mexico Chlorella chloroplast) constructed with endogenous recombination regions, namely, 16S–trn I (left) and trn A–23S (right), and the Prrn promoter. Human basic fibroblast growth factor (bFGF) was adopted as a target biopharmaceutical to establish the chloroplast expression approach. The plasmid pCMCC:bFGF was transformed into C. vulgaris via electroporation using simple carbohydrate-based buffers, which aided in the transfer of the transgene into the chloroplast genome. Cells transformed with the pCMCC:bFGF vector were selected using kanamycin, and resistant colonies were analyzed by polymerase chain reaction and Western blotting to confirm the presence of the transgene and the recombinant bFGF, respectively. The bFGF that accumulated in the transplastomic C. vulgaris clones ranged from 0.26 to 1.42 ng/g fresh weight of biomass, and it was quantified by enzyme-linked immunosorbent assay. Therefore, the designed expression vector, in combination with an optimized electroporation protocol, constitutes a viable approach to successfully develop transplastomic lines of C. vulgaris for the potential low-cost production of biopharmaceuticals using this algal species. This study paves the way for the establishment of chloroplast biotechnology in microalgae other than the model organism C. reinhardtii.

show abstract

Paving the way for the production of secretory proteins by yeast cell factories

Cited by 14 publications

References 11 publications

Recent Developments in Bioprocessing of Recombinant Proteins: Expression Hosts and Process Development

Recent Developments in Bioprocessing of Recombinant Proteins: Expression Hosts and Process Development

Mammalian cell lines used in bioprocessing

Construction and Validation of a Chloroplast Expression Vector for the Production of Recombinant Proteins in Chlorella vulgaris

Contact Info

Product

Resources

About