Sheng wu gong cheng xue bao = Chinese journal of biotechnology最新文献

Itaconate is a pivotal intermediate metabolite in the tricarboxylic acid (TCA) cycle of immune cells. It is produced by decarboxylation of cis-aconitic acid under the catalysis of aconitate decarboxylase 1 (ACOD1), which is encoded by the immune response gene 1 (IRG1). Itaconate has become a focal point of research on immunometabolism. Studies have demonstrated that itaconate plays a crucial role in diseases by regulating inflammation, remodeling cell metabolism, and participating in epigenetic regulation. This paper reviewed the research progress in itaconnate from its chemical structure, regulatory effects on different diseases, and mechanisms, proposes the future research directions, aiming to provide a theoretical basis for the development of itaconate-related drugs.

This study aims to establish a time-resolved fluorescence immunochromatography method for simultaneous determination of human papillomavirus (HPV) type 16 E6 and L1 protein concentrations. The amount of lanthanide microsphere-labeled antibodies, the concentration of coated antibodies, and the reaction time were optimized, and then a test strip for the simultaneous determination of the protein concentrations was prepared. The performance of the detection method was evaluated based on the concordance of the results from clinical practice. The optimal conditions were 8 μg and 10 μg of HPV16 L1 and E6-labeled antibodies, respectively, 1.5 mg/mL coated antibodies, and reaction for 10 min. The detection with the established method for L1 and E6 proteins showed the linear ranges of 5-320 ng/mL and 2-64 ng/mL and the lowest limits of detection of 1.78 ng/mL and 1.09 ng/mL, respectively. There was no cross reaction with human immunodeficiency virus (HIV), treponema pallidum (TP), or HPV18 E6 and L1 proteins. The average recovery rate of the established method was between 97% and 107%. The test strip prepared in this study showed the sensitivity, specificity, and diagnostic accuracy of 97.46%, 90.57%, and 95.32%, respectively, in distinguishing patients with cervical cancer and precancerous lesions from healthy subjects, with the area under the curve (AUC) of 0.980 1 and 95% Confidence Interval (CI) of 0.956 5 to 1.000 0. The time-resolved fluorescence immunochromatography combined with the test strips prepared in this study showed high sensitivity, high accuracy, simple operation, and rapid reaction in the quantitation of HPV16 E6 and L1 proteins. It thus can be used as an auxiliary method for the diagnosis and early screening of cervical cancer and precancerous lesions and the assessment of disease course.

To investigate the mechanism of the major capsid protein VP5 (encoded by the UL19 gene) of oncolytic herpes simplex virus type Ⅱ (oHSV2) in regulating the antitumor function of immune cells, we constructed a mouse breast cancer cell line 4T1-iRFP-VP5-GFP stably expressing VP5 protein, near-infrared fluorescent protein (iRFP), and green fluorescent protein (GFP) by using the PiggyBac transposon system. Flow cytometry and Western blotting were employed to screen the monoclonal cell lines expressing both GFP and VP5 and examine the expression stability of UL19 in the constructed cell line. The results of SYBR Green I real-time PCR and Western blotting showed that the copies of UL19 and the expression level of VP5 protein in the 15th passage of 4T1-iRFP-VP5-GFP cells were significantly higher than those in the 4T1 cells transiently transfected with UL19, demonstrating the stable insertion of UL19 into the 4T1 cell genome. The real-time cell analysis (RTCA) was employed to monitor the proliferation of 4T1-iRFP-VP5-GFP cells, which showed similar proliferation activity to their parental 4T1 cells. Further studies confirmed that NK92 cells exhibited stronger cytotoxicity against 4T1-iRFP-VP5-GFP cells than against 4T1 cells. This study layed a foundation for elucidating the role of VP5 protein in regulating immune cells, including T cells and NK cells, via HLA-E in 4T1 cells to exert the anti-tumor function.

Some bacteria can selectively colonize the tumor site and inhibit tumor growth, serving as ideal vehicles for delivering antitumor drugs. The system of delivering antitumor drugs with live bacteria as vehicles is characterized by good biocompatibility and precise targeting. However, the development of bacteria as drug delivery vehicles is limited by their own immunogenicity. In this paper, the selection of chassis bacteria, bacterial loading drug strategies, antitumor drug delivery applications and their limitations are elaborated in detail, and its future development direction is envisioned, with a view to providing a reference for the study of live bacteria as antitumor drug delivery carriers.

Bispecific antibody drugs have enhanced efficacy and safety compared with therapeutic monoclonal antibody drugs, gradually becoming a new generation of effective therapies. With the development of genetic engineering and the maturity of the industry, increasing studies have been carried out on bispecific and multi-specific antibody drugs, and the application indications of these drugs are expanding, which lays a foundation for satisfying the clinical needs and creating clinical values. This paper provides a comprehensive review of the development stages, indications, target combinations, structural formats, and mechanisms of bispecific and multi-specific antibody drugs. It also discusses key points and strategies for differentiated drug discovery, facilitating the rapid development of innovative therapies, providing theoretical support for the clinical translation of these drugs, and offering more precise and effective treatment options for clinical practice.

Foodborne pathogens are one of the major factors leading to food safety issues, causing harm to the national economy and people's livelihood. Achieving rapid detection of foodborne pathogens is currently a key strategy for preventing and controlling foodborne diseases. Antibodies naturally possess high specificity and sensitivity, serving as preferred tools for specific recognition of foodborne pathogens. We list the main methods for detecting foodborne pathogens, introduce the evolution and development of polyclonal antibodies, monoclonal antibodies, and genetically engineered antibodies, and review the application of different antibody technologies in the rapid detection of foodborne pathogens. Furthermore, we recognize that the combination of antibody technology and other foodborne pathogen detection methods is currently a reliable means to improve detection performance. Finally, we elaborate on the existing limitations of different antibodies and summarize the current research status and potential issues, aiming to provide a theoretical basis and practical ideas for the development of rapid detection of foodborne pathogens.

Vaccination is the most effective measure for reducing and preventing influenza and related complications. In this study, we analyzed the mutation trend and the antigen dominant site changes of the amino acid sequence of hemagglutinin subunit 1 (HA1) of human influenza A virus (IAV) in the northern hemisphere from 2012 to 2022. According to the HA1 sequences of A/Darwin/6/2021 (H3N2) and A/Wisconsin/588/2019 (H1N1) recommended by the World Health Organization in the 2022 influenza season in northern hemisphere, we employed the mosaic algorithm to design three Mosaic-HA1 antigens through stepwise substitution. Mosaic-HA1 was expressed and purified in 293F cells and then mixed with the alum adjuvant at a volume ratio of 1:1. The mixture was used to immunize BALB/c mice, and the immunogenicity was evaluated. Enzyme-linked immunosorbent assay showed that Mosaic-HA1 induced the production of IgG targeting two types of HA1, the specific IgG titers for binding to H3 protein and H1 protein reached 10⁵ and 10³ respectively. The challenge test showed that Mosaic-HA1 protected mice from H3N2 or H1N1. This study designs the vaccines by recombination of major antigenic sites in different subtypes of IAV, giving new insights into the development of multivalent subunit vaccines against influenza.

With the advancement of synthetic biology, recombinant proteins are poised to play a significant role in medical applications. The scaled manufacturing is a pillar for the extensive application and development of recombinant proteins across various fields. In the large-scale production process of recombinant proteins, the removal and detection of endotoxins are essential to reduce their levels to safe thresholds in the final products. Currently, establishing stringent endotoxin limits for different recombinant protein products is a crucial aspect of safety assessment. This review begins by shedding light on the pathogenicity of endotoxins and discusses the methods for the removal and determination of endotoxins during the production processes of recombinant proteins. Subsequently, this review summarizes the endotoxin limits in industries such as biologics, medical devices, and human recombinant DNA products, particularly those in recombinant protein injection products. It is highlighted that regardless of whether the hosts for recombinant protein expression are bacteria or not, endotoxin testing is required for the final products of injectable recombinant proteins, and compliance with relevant industry standards is necessary. This review aims to provide a reference for the research on endotoxins in the large-scale production process of recombinant proteins.

Conventional administration methods have problems including low bioavailability, complex operation process, and discomfort of patients with fear of needles. Transdermal delivery can avoid these problems, whereas most drugs are difficult to directly penetrate the skin cuticle and reach the diseased site. Microneedling is an emerging method of local drug delivery, enabling the drug penetration through the stratum corneum of the skin in a minimally invasive manner and delivering the drug directly to the diseased site, thereby improving the treatment effect. Stimuli-responsive microneedles have attracted much attention because of the spatiotemporal controllability, high drug delivery efficiency, and mild potential side effects. This review introduced the commonly used materials and various types of stimuli-responsive microneedles and the drug release mechanisms. In addition, this paper expounded the biomedical applications of stimuli-responsive microneedles as drug delivery systems in response to different stimuli and discusses the challenges and potential solutions for stimuli-responsive microneedles.

Compared with conventional vaccines, mRNA vaccines have considerable advantages in design, production, and application, especially in dealing with emerging infectious diseases. Particularly, mRNA vaccines were the first to be recommended by the World Health Organization for emergency use during the COVID-19 pandemic. A key to the design of mRNA vaccines is to ensure the stable and sufficient expression of the encoded protein in the recipient. In recent years, advances have been attained in the experimental and computational research in this area. This review focused on the progress and problems in improving the translation efficiency of mRNA vaccines in recent years, aiming to promote related research.