Role of Engineered Proteins as Therapeutic Formulations
Peter Grunwald in Pharmaceutical Biocatalysis, 2019
Non-homologous recombination methods have also been developed to produce chimeric proteins via recombination of parental sequences which are non-homologous. Non-homologous parental sequences imply that they share very little sequence identity. Such non-homologous sequences cannot be recombined using the above-mentioned homologous recombination methods. Exon shuffling is the one of the non-homologous recombination methods developed by Kolkman and Stemmer (Kolkman and Stemmer, 2001). This method essentially requires mixing of exons from different protein genes via recombination at the intervening regions known as introns. Chimeric protein will exhibit the features of all the recombining exons (Kolkman and Stemmer, 2001). Further, diversity can be inculcated in these chimeras by insertion, deletion, or substitution during the recombination event. Few other non-homologous recombination techniques such as incremental truncation for the creation of hybrid enzymes (ITCHY) (Ostermeier et al., 1999), sequence homology independent protein recombination (SHIPREC) (Sieber et al., 2001), Degenerate Homo-duplex Recombination (DHR) (Coco et al., 2002), Random Multi-recombinant PCR (RM-PCR) (Tsuji et al., 2001), golden gate shuffling (GGS) Recombination (Engler et al., 2009), Integron (Bikard et al., 2010), Y-ligation-based shuffling (YLBS) (Kitamura et al., 2002) have also been developed to generate chimeric proteins with novel functionalities.
Order Blubervirales: Surface Protein
Paul Pumpens, Peter Pushko, Philippe Le Mercier in Virus-Like Particles, 2022
Thus, first, Rutgers et al. (1988) generated two variants of the fusion proteins, where 1 or 15 tetrapeptide NANP repeats from the circumsporozoite protein (CSP) of P. falciparum were added before the aa position 42 of the preS2 part of the MHBs protein. In both hybrid proteins one tetrapeptide repeat NVDP was also present resulting from the fusion of the CSP sequences NV to the preS2 sequences DP. The synthesis of the chimeric proteins and generation of the 22-nm-like mosaic particles, which were formed by the chimeras together with the natural SHBs protein translated from its intrinsic ATG codon, was achieved in S. cerevisiae. The initial construct with the 15 NANP and one NVDP, referred to as R16-HBsAg, was formulated with alum and proved safe and immunogenic in 20 hepatitis B-seronegative adults at the University of Nijmegen (Vreden et al. 1991).
Molecular Diagnosis of Pleural Neoplasms
Philip T. Cagle, Timothy C. Allen, Mary Beth Beasley in Diagnostic Pulmonary Pathology, 2008
The use of molecular studies to stratify patients into prognostic categories remains a focus of research. In a meta-analysis conducted in 2003, EWS-FLI1 type 1 fusion transcript containing tumors were associated with improved disease-free survival (61). The suggestion that different chimeric proteins have varied impact on cellular growth leads to an expectation that future studies will result in prognosis stratification on the basis of type of fusion transcript. This may favor a molecular technique that allows for detection of the specific fusion transcript type such as qRT-PCR. This technique also provides the potential for quantitation. While fusion transcript detection in blood (62) and bone marrow (63) is possible, it is not clear that identification of fusion transcripts in serum or bone marrow predicts tumor progression (64); such identification in longer-term survivors may predict recurrence (65).
The potential of plant-made vaccines to fight picornavirus
Published in Expert Review of Vaccines, 2020
Omayra C. Bolaños-Martínez, Sergio Rosales-Mendoza
Plant-based vaccines are being adopted as an alternative to generate new HAV vaccines. In 2011, Chung et al. reported the expression and immunogenicity of a recombinant chimeric protein (HAV VP1-Fc) containing the VP1 viral protein and an Fc antibody fragment. The chimeric protein was detected and subsequently purified. The expression levels reached up to 0.6% TSP. The immunogenicity was assessed in mice after i.p. immunization (4 times at 2-week intervals with 30 µg of VP1). Strong IgG response to VP1-Fc in sera was obtained from the group immunized with the plant-made antigen. Furthermore, splenocytes from immunized mice showed increased INF-γ and IL-4 production upon antigenic stimuli. These results indicated the induction of Th1 and Th2-type cytokines [42]. The assessment of the protective capacity of this vaccine in appropriate animal models will provide solid evidence of its true potential.
Molecular features, prognosis, and novel treatment options for pediatric acute megakaryoblastic leukemia
Published in Expert Review of Hematology, 2019
Federico De Marchi, Marito Araki, Norio Komatsu
As aforementioned, CBFA2T3–GLIS2 is one of the recurrent translocations in non-DS AMegL. The chimeric protein binds to DNA to control transcription factors, resulting in self-renewal, proliferation, and differentiation blockage. It has been demonstrated that specific interference with this fusion protein could promote megakaryocytic maturation and proliferative inhibition [42]. In this aspect, aurora kinase A is a negative regulator of polyploidization, and its selective inhibitor MLN8237 (alisertib) induces megakaryocytic maturation in AMegL [72,73]. CBFA2T3–GLIS2 is also associated with the overexpression of Hedgehog-related genes, with GLIS2 being homologous to the final effectors of the pathway. Overexpression of these genes is a known negative prognostic factor in other AML subtypes and is, thus, considered as a therapeutic target [74]. A recent study tested GANT61, the strongest GLI inhibitor, and found that cells harboring CBFA2T3–GLIS2 underwent apoptosis and G1 cell cycle arrest [75].
Enhancement of antibody-dependent cellular cytotoxicity of cetuximab by a chimeric protein encompassing interleukin-15
Published in OncoImmunology, 2018
Maria Carmen Ochoa, Luna Minute, Ascensión López, Elisabeth Pérez-Ruiz, Celia Gomar, Marcos Vasquez, Susana Inoges, Iñaki Etxeberria, Inmaculada Rodriguez, Saray Garasa, Jan-Peter Andreas Mayer, Peter Wirtz, Ignacio Melero, Pedro Berraondo
An increase in ADCC mediated by tumor-targeted monoclonal antibodies upon combination with different IL15 variants has been recently reported.12,13 Our results with the chimeric Sushi-IL15-Apo fully support these previous findings. The novel chimeric protein enhanced the number of NK cells due to an increase in survival and proliferation. In addition, the chimeric protein increased the cytotoxic potential on a per cell basis. Thus, the combination of increased numbers of effector cells and their cytotoxic performance resulted in enhanced ADCC as mediated by cetuximab. This effect was not only demonstrated in vitro, but also observed in immunodeficient mice xenografted with EGFR+ HT-29 tumor cells in which adoptive transfer of PBMCs in combination with cetuximab and the Sushi-IL15-Apo attained maximal tumor cell elimination, and in a syngeneic model of a subcutaneous MC38 colon cancer expressing EGFR.
Related Knowledge Centers
- Chimera
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- Protein Domain
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