Article
Immunology
Nicholas J. MacDonald, Kavita Singh, Karine Reiter, Nguyen Vu, Richard Shimp, Apostolos G. Gittis, Beth Chen, Martin Burkhardt, Baoshan Zhang, Zhixiong Wang, Raul Herrera, Mackenzie Moler, Duck-Yeon Lee, Sachy Orr-Gonzalez, Jessica Herrod, Lynn E. Lambert, Kelly M. Rausch, Olga Muratova, David S. Jones, Yimin Wu, Albert J. Jin, David N. Garboczi, Patrick E. Duffy, David L. Narum
Summary: The development of a malaria vaccine that can block transmission of the parasite from mosquitoes to humans is crucial for malaria elimination efforts. The use of Pfs25, a protein on the surface of the parasite, is being investigated as a transmission-blocking vaccine. Two recombinant forms of Pfs25, Pfs25H and Pfs25M, have been studied for their structural and immunological differences.
Article
Engineering, Biomedical
Shuo-Fu Yuan, Sierra M. Brooks, Annalee W. Nguyen, Wen-Ling Lin, Trevor G. Johnston, Jennifer A. Maynard, Alshakim Nelson, Hal S. Alper
Summary: Traditional production of industrial and therapeutic proteins by eukaryotic cells typically requires large-scale fermentation capacity, which is not easily portable or reusable for on-demand protein production applications. In this study, a hydrogel-based technique was used to immobilize engineered Pichia pastoris for preservable, on-demand production and secretion of medium- and high-molecular weight proteins. This platform shows potential for establishing a cost-effective bioprocessing strategy for on-demand protein production.
BIOACTIVE MATERIALS
(2021)
Article
Biotechnology & Applied Microbiology
Chatchol Kongsinkaew, Kittisak Ajariyakhajorn, Viroj Boonyaratanakornkit, Sarintip Sooksai, Soisuda Pornpukdeewattana, Warawut Krusong, Azis Boing Sitanggang, Theppanya Charoenrat
Summary: During recombinant protein expression in Pichia pastoris under the alcohol oxidase promoter, the culture supernatant developed green and yellow pigments. We designed a sequential cross-flow filtration method using microfiltration and ultrafiltration membranes to eliminate impurities and reduce the pigments. The method effectively removed host cell proteins and reduced the green pigment, while allowing the target peptide to be recovered.
FOOD AND BIOPRODUCTS PROCESSING
(2023)
Article
Multidisciplinary Sciences
Parag Goyal, Harish Venkatraman Pai, Phanichand Kodali, Bhavesh Vats, Navratna Vajpai, Shankara Annegowda, Krishnappa Mane, Shamini Mohan, Shruti Saxena, Anil Bangalore Veerabhadraia, Milee Palande, Preethy Sasankan Nair, Digvijay Chandrashekar More, Umamaheshwara Rao Karudumpa, Kunala Jyothirmai, Adroha Bhattacharya, Frida Almeida, Santosh Gulab Khyade, Shankara Gouda, Daniel J. Ranayhossaini, Praveen Reddy Moole, Jeffrey P. Smith, Abhijit Barve, Ramakrishnan Melarkode, Rajesh Ullanat
Summary: MYL-1501D is a biosimilar insulin glargine designed to offer patients greater access to insulin glargine with comparable efficacy and safety to marketed products. Comprehensive studies have shown high similarity in structural and functional properties between MYL-1501D and marketed insulin glargine products, suggesting potential as a cost-effective alternative treatment option.
Review
Biotechnology & Applied Microbiology
Antonio Carlesso, Raquel Delgado, Oriol Ruiz Isant, Owens Uwangue, Dylan Valli, Roslyn M. Bill, Kristina Hedfalk
Summary: This study emphasizes the importance of yeast as a production host for eukaryotic membrane proteins. Yeast remains a useful host for studying α-helical membrane proteins, with both homologous and heterologous production being common in different yeast hosts.
FEMS YEAST RESEARCH
(2022)
Article
Multidisciplinary Sciences
Neil C. Dalvie, Sergio A. Rodriguez-Aponte, Brittany L. Hartwell, Lisa H. Tostanoski, Andrew M. Biedermann, Laura E. Crowell, Kawaljit Kaur, Ozan S. Kumru, Lauren Carter, Jingyou Yu, Aiquan Chang, Katherine McMahan, Thomas Courant, Celia Lebas, Ashley A. Lemnios, Kristen A. Rodrigues, Murillo Silva, Ryan S. Johnston, Christopher A. Naranjo, Mary Kate Tracey, Joseph R. Brady, Charles A. Whittaker, Dongsoo Yun, Natalie Brunette, Jing Yang Wang, Carl Walkey, Brooke Fiala, Swagata Kar, Maciel Porto, Megan Lok, Hanne Andersen, Mark G. Lewis, Kerry R. Love, Danielle L. Camp, Judith Maxwell Silverman, Harry Kleanthous, Sangeeta B. Joshi, David B. Volkin, Patrice M. Dubois, Nicolas Collin, Neil P. King, Dan H. Barouch, Darrell J. Irvine, J. Christopher Love
Summary: Subunit vaccines based on recombinant proteins are crucial tools for controlling COVID-19 due to their low manufacturing cost, scalability, and proven safety and efficacy. Engineered RBD variants exhibit high manufacturability and immunogenicity, showing enhanced immune responses after a single dose and cross-reactivity to new variants of concern.
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
(2021)
Review
Biotechnology & Applied Microbiology
Chao Liu, Jin-Song Gong, Chang Su, Hui Li, Heng Li, Zhi-Ming Rao, Zheng-Hong Xu, Jin-Song Shi
Summary: Pichia pastoris has been recognized as an important platform for heterologous protein production. This article summarizes the process of protein secretion, modification, and transportation in P. pastoris, as well as the roles played by key proteins and their co-expression effects.
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
(2022)
Article
Biotechnology & Applied Microbiology
Jungsoon Lee, Zhuyun Liu, Wen-Hsiang Chen, Junfei Wei, Rakhi Kundu, Rakesh Adhikari, Joanne Altieri Rivera, Portia M. Gillespie, Ulrich Strych, Bin Zhan, Peter J. Hotez, Maria Elena Bottazzi
Summary: The study presented the development and scale-up optimization of production and purification process for RBD219-N1C1 protein antigen against SARS-CoV-2, with the selected process yielding high purity and recovery rate of the product. Different chromatographic purification methods were compared, and the best process was chosen based on analytical studies. An optimized production process was proposed, which can generate high-quality RBD219-N1C1 protein antigen at a low cost and scalable level.
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
(2021)
Article
Food Science & Technology
Jewel Ann Joseph, Simen Akkermans, Jan F. M. Van Impe
Summary: The sweet protein thaumatin is a promising sugar replacer, and its large-scale extraction from natural plants can be limited in terms of raw material availability and production costs. This study evaluated the impact of pH and temperature on the production of thaumatin II protein using Pichia pastoris, and found that a pH of 6.0 and a temperature of 30 degrees C resulted in the highest protein levels.
Article
Biotechnology & Applied Microbiology
Troy R. Alva, Melanie Riera, Justin W. Chartron
Summary: By analyzing the translatomes of Komagataella phaffii, it was found that cell-wall components represent the greatest number of nascent chains entering the ER during exponential growth in rich media. Transcripts encoding the transmembrane protein PMA1 sequester more ribosomes at the ER membrane than any others. Given the bottlenecks in heterologous protein production, certain host proteins may be potential targets for pathway improvement.
MICROBIAL CELL FACTORIES
(2021)
Article
Biology
Binyang Deng, Yu Yue, Jun Yang, Mingjun Yang, Qiong Xing, Hang Peng, Fei Wang, Ming Li, Lixin Ma, Chao Zhai
Summary: In this study, highly glycosylated IsPETase enzyme was obtained by controlling the glycosylation modification of PETase in Pichia pastoris, improving the enzyme's specific activity and thermostability. The partially deglycosylated IsPETase still maintained high thermostability and could completely degrade PET flakes at 50 degrees C. This study provides a strategy to modulate the thermostable IsPETase through glycosylation engineering, promoting PET biodegradation.
COMMUNICATIONS BIOLOGY
(2023)
Article
Biotechnology & Applied Microbiology
Fidan Erden-Karaoglan, Mert Karaoglan
Summary: This study investigated the applicability of heterologous yeast promoters in Pichia pastoris, and found inducible and constitutive regulation modes as well as the optimal carbon sources for these promoters. The study also determined the DNA region with the highest activity for all promoters through promoter deletion analysis.
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
(2022)
Article
Biochemistry & Molecular Biology
Khadijah A. Altammar, Jonathan Guyang Ling, Hussein M. Al-Bajalan, Iuan-Sheau Chin, Mukram Mohamed Mackeen, Nor Muhammad Mahadi, Abdul Munir Abdul Murad, Farah Diba Abu Bakar
Summary: Cutinases are hydrolytic enzymes that can degrade plant cutin. A new cutinase homolog, AnCUT3, has been identified and characterized in this study. The recombinant AnCUT3 exhibited optimal temperature and pH, as well as high catalytic efficiency. It also showed potential for degrading and modifying the surface of certain polymers.
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
(2022)
Article
Biotechnology & Applied Microbiology
Jiao Meng, Shufan Liu, Le Gao, Kai Hong, Shuguang Liu, Xin Wu
Summary: This study elucidated a unique mechanism of P. pastoris for efficient methanol utilization, higher temperature resistance, and high protein synthesis, providing a P. pastoris cell factory for SCP production with environmental, economic, and nutritional benefits. Through adaptive laboratory evolution and metabolic engineering, the engineered strain produced high levels of SCP from methanol in a pilot-scale fed-batch culture, highlighting its potential in sustainable biomanufacturing.
MICROBIAL CELL FACTORIES
(2023)
Article
Food Science & Technology
Xue Zhang, Chunyue Zhang, Mian Zhou, Quanming Xia, Liqiang Fan, Liming Zhao
Summary: Chitin and its derivatives have diverse applications, and this research achieved increased chitin content in yeast cell walls through gene overexpression and culture condition optimization. This is the first report of chitin production in engineered Pichia pastoris via biosynthetic pathway enhancement.
Article
Biochemical Research Methods
Juri Sakata, Toshifumi Tatsumi, Akira Sugiyama, Akihiro Shimizu, Yuya Inagaki, Hiroto Katoh, Takefumi Yamashita, Kazuki Takahashi, Sho Aki, Yudai Kaneko, Takeshi Kawamura, Mai Miura, Masazumi Ishii, Tsuyoshi Osawa, Toshiya Tanaka, Shumpei Ishikawa, Masanobu Tsukagoshi, Michael Chansler, Tatsuhiko Kodama, Motomu Kanai, Hidetoshi Tokuyama, Kenzo Yamatsugu
Summary: This study reports an expression, refolding, and purification method for antibody-mimetic drug conjugate (AMDC). The AMDC uses a variable heavy chain of heavy chain-only antibodies (VHHs) as the targeting protein and non-covalently binds to a potent DNA-alkylating drug. The AMDC shows strong cytotoxic effects on cancer cells.
PROTEIN EXPRESSION AND PURIFICATION
(2024)
Article
Biochemical Research Methods
Yao Wang, Shaoting Weng, Yajie Tang, Sen Lin, Xiayue Liu, Wenhui Zhang, Gang Liu, Boomi Pandi, Yinrong Wu, Lei Ma, Lin Wang
Summary: In this study, a high yield of recombinant CD20 and claudin 18.2 proteins was achieved using an in vitro coupled transcription-translation system. The results showed that rituximab has a high affinity with CD20 protein. This study provides a novel concept for promoting the expression of multi-pass transmembrane proteins and lays the foundation for large-scale industrial production of membrane-associated drug targets.
PROTEIN EXPRESSION AND PURIFICATION
(2024)
Article
Biochemical Research Methods
Xiaobing Chen, Zijuan Zou, Wei Li, Xu Dong, Yi Chen, Yan Lu, Mingyue Zhu, Mengsen Li, Bo Lin
Summary: In this study, the fusion protein ImI-AFP3, composed of alpha-Conotoxin ImI and human alpha fetoprotein domain 3 (AFP3), was found to inhibit the growth and migration of lung cancer cells and showed synergistic effects with the drug gefitinib. These findings suggest that ImI-AFP3 is a promising candidate for the development of anticancer drugs.
PROTEIN EXPRESSION AND PURIFICATION
(2024)
Article
Biochemical Research Methods
Angel Castillo-Corujo, Mirva J. Saaranen, Lloyd W. Ruddock
Summary: This study successfully expressed two Fabs antibodies in the cytoplasm of E. coli using the CyDisCo system, achieving high yields and biological activity under industrially relevant fermentation conditions.
PROTEIN EXPRESSION AND PURIFICATION
(2024)
Article
Biochemical Research Methods
Junling Guo, Zhongyi Cheng, Zhemin Zhou
Summary: This study focused on a halophilic archaeal nitrile hydratase (NHase) and found that it exhibited higher tolerance to substrates and products compared to NHases from other sources. The unique genetic structure of this highly stable archaeal NHase could provide a theoretical foundation for modifying and enhancing the industrial application of NHase.
PROTEIN EXPRESSION AND PURIFICATION
(2024)
Article
Biochemical Research Methods
Yunlong Shen, Ruirui Zhang, Xiaohua Jiang, Jinliang Yang
Summary: The study developed a monoclonal antibody, B1M023, that can bind to LILRB1 with high affinity and block its binding to HLA-G. This antibody can promote the activation and IFN-gamma secretion of T cells, suggesting its potential applications in concomitant diagnosis and tumor immunotherapy.
PROTEIN EXPRESSION AND PURIFICATION
(2024)
Article
Biochemical Research Methods
Jun Jiang, Zhengqiang Jiang, Qiaojuan Yan, Susu Han, Shaoqing Yang
Summary: A novel alginate lyase gene from a marine bacterium showed efficient enzyme activity and stability, producing high conversion ratios of alginate oligosaccharides.
PROTEIN EXPRESSION AND PURIFICATION
(2024)
Article
Biochemical Research Methods
Xianghua Xiong, Yujin Qiu, Jiahao Zheng, Ling Zhou, Qingyang Wang, Jinglun Pang, Weicai Zhang, Huipeng Chen, Gang Liu, Xiaodong Han
Summary: A specific monoclonal antibody ML419 has been found to disrupt the recognition between Botulinum neurotoxin serotype A (BoNT/A) and FGFR3, effectively preventing BoNT/A from entering neurons. In vivo experiments show that ML419 has a strong protective effect, making it a promising candidate for the development of therapeutics against BoNT/A.
PROTEIN EXPRESSION AND PURIFICATION
(2024)
Article
Biochemical Research Methods
Maria Rain Jennings, Soohyon Min, Grace S. Xu, Kassandra Homayuni, Bhavana Suresh, Yusef Amir Haikal, John Blazeck
Summary: This study successfully produced Homo sapiens adenosine deaminase isoform 1 (HsADA1) and its variants through optimizing the recombinant expression process. The D8N variant of HsADA1 was found to be about 30% less active than the wildtype, but it better retained its activity in human serum. Additionally, the study revealed a previously undescribed phenomenon involving albumin that contributed to the increased activity of HsADA1 and the D8N variant in serum.
PROTEIN EXPRESSION AND PURIFICATION
(2024)
Article
Biochemical Research Methods
Oinam Sangita Devi, Senjam Sunil Singh, K. Rana, Sorokhaibam Jibankumar Singh, Wayenbam Sobhachandra Singh
Summary: A new lectin with hemagglutination activity was purified from the rhizome of Xanthosoma violaceum Schott. The lectin showed different reactions towards human red blood cells of different blood groups. It exhibited optimal hemagglutination activity at specific temperature and pH range, and showed good stability.
PROTEIN EXPRESSION AND PURIFICATION
(2024)