Potential of Antibody Therapy for Respiratory Virus Infections
Sunit K. Singh in Human Respiratory Viral Infections, 2014
Most yeast display technologies are based on Saccharomyces cerevisiae, however. Pichia pastoris has been introduced as an alternative species as it does not revert to anaerobic metabolism under oxygen deprivation conditions, and thus does not produce toxic metabolites. It is also easier to genetically modify and express, and is stable in both haploid and diploid forms [75]. However, the posttranslational glycosylation in P. pastoris differs greatly from humans and could result in immunogenicity if these mAbs are used for human therapy. Methods to fine-tune glycosylation of antibodies in these systems, such as the removal of genes encoding certain mannose sugars in P. pastoris pathways [76], are currently being studied.
Recombinant Antibodies
Siegfried Matzku, Rolf A. Stahel in Antibodies in Diagnosis and Therapy, 2019
The methylotrophic yeast Pichia pastoris has been shown to be suitable for the high-level expression of various heterologous proteins, either intracellular or secreted into the culture supernatant (see review by Cregg et al., 1993). Recently, Ridder et al. (1995) demonstrated the applicability of the Pichia expression system for the secretion of antibody fragments. A rabbit scFv against the human leukemia inhibitory factor (hLIF) was expressed in Pichia cells using the expression vector pPIC9 that provides the α-mating factor signal sequence for secretion and the HIS4 gene for selection of the recombinant yeast clones. The yield was about 100 mg per liter of shake flask culture.
Order Zurhausenvirales
Paul Pumpens, Peter Pushko, Philippe Le Mercier in Virus-Like Particles, 2022
The successful story of the yeast-supported production of the HPV VLPs started in early 1990s when Sasagawa et al. (1995) demonstrated the self-assembly of the HPV6 and HPV16 VLPs from the expressed L1 alone and both L1 and L2 together, in Schizosaccharomyces pombe, although L2 was not incorporated into the VLPs. Remarkably, the two HPV16 L1 variants, isolated from benign cervical samples, produced many more (68- and 14-fold) VLPs than the prototype L1 derived from cervical carcinoma. A bit later, the HPV6a L1 or the L1 together with L2 ORFs were expressed in the yeast Saccharomyces cerevisiae, and the self-assembly of the L1 protein into the VLPs was demonstrated both in the L1 as well as L1 + L2 coexpressing yeast strains (Hofmann et al. 1995). In this case the copurification of the L1 and L2 proteins showed complex formation of the L1 and L2 proteins in the yeast-derived VLPs of the coexpressing strains. Next, the S. cerevisiae yeast was used to get VLPs by the expression of the HPV6/11 hybrid L1 (Neeper et al. 1996; Cook et al. 1999; Brown et al. 2001), HPV16 L1 (Koutsky et al. 2002; Kim SN et al. 2007; Park et al. 2008; Kim HJ et al. 2012), and HPV18 L1 (Woo et al. 2008) genes. Bazan et al. (2009) performed the expression of the codon-optimized HPV16 L1 gene and characterization of the corresponding VLPs produced by the methylotrophic yeast Pichia pastoris. The expression of the codon-optimized L1 genes of HPV16 and HPV18 in Pichia pastoris was presented by Hanumantha Rao et al. (2011). Furthermore, the L1 genes of HPV16, 18, and 33 (Coimbra et al. 2011) and HPV58 (Jiang et al. 2011) were expressed in Pichia pastoris.
Effects of the Cobalt-60 gamma radiation on Pichia pastoris glyceraldehyde-3-phosphate dehydrogenase
Published in International Journal of Radiation Biology, 2022
Abdelghani Iddar, Mohammed El Mzibri, Adnane Moutaouakkil
This work was carried out to assess the adaptive response of Pichia pastoris, used as a eukaryotic model, after gamma irradiation. Indeed, the methylotrophic yeast P. pastoris represents a good eukaryotic model for many studies. Similar to Escherichia coli, P. pastoris can be grown easily in a laboratory with a relatively short regeneration time and it is able to grow up to high cell densities, resulting in high protein production yields (Potvin et al. 2010). P. pastoris is a single cell organism considered as most effective heterologous expression system with an easy carry out of genetic manipulation and it has become more extensively used for industrial applications (Gasser et al. 2013). P. pastoris allowed the expression and production of proteins that could not be produced efficiently by bacteria or other host organisms (Cereghino et al. 2002). Moreover, P. pastoris has a number of promoters that allow the regulation of proteins in the cell (Stadlmayr et al. 2010). The purpose of this study was to evaluate the effect of Cobalt-60 gamma radiation on P. pastoris GAPDH activity and protein levels. The effect of radiation on induction of some oxidative damages and on antioxidant enzymes activities was also explored.
Complement inhibitor factor H expressed by breast cancer cells differentiates CD14+ human monocytes into immunosuppressive macrophages
Published in OncoImmunology, 2020
Karolina I. Smolag, Christine M. Mueni, Karin Leandersson, Karin Jirström, Catharina Hagerling, Matthias Mörgelin, Paul N. Barlow, Myriam Martin, Anna M. Blom
FH20 and α1-AT21 were purified from human plasma (at least 95% pure). LPS contamination was tested using a Limulus amebocyte lysate Endochrome-K assay (#R1708K; Charles River) and levels have been determined to <0.01–0.03 EU/ml in working concentration for FH and <0.04 for α1-AT. FH was incubated for 30 min at 96°C to obtain D-FH.22 FH CCP1-7 and CCP19-20 were produced in Pichia pastoris.23 Nucleosomes were isolated with Nucleosome preparation kit (#53504; Active Motif) and labeled in parallel with Jurkat T-cells with pHrodo ester (#P35369; Invitrogen).
Virus-like particle-based nanocarriers as an emerging platform for drug delivery
Published in Journal of Drug Targeting, 2023
Bingchuan Yuan, Yang Liu, Meilin Lv, Yilei Sui, Shenghua Hou, Tinghui Yang, Zakia Belhadj, Yulong Zhou, Naidan Chang, Yachao Ren, Changhao Sun
Pichia pastoris and Hansenula are common yeast cells. Unlike bacterial expression system, the genes encoding VLP assembly proteins are transferred to yeast cells using yeast shuttle vectors. The expressed proteins to be assembled can be modified, such as by glycosylation and phosphorylation [134], after translation in the yeast expression system. Yeast cells may also express a variety of subunit proteins and assemble them into VLPs. For example, three rotavirus structural proteins can be simultaneously expressed and assembled into multiple VLPs [135].
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