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( Aqsa Ahmed ),( Abdullah Ahmad ),( Renhan Li ),( Waleed Al-ansi ),( Momal Fatima ),( Bilal Sajid Mushtaq ),( Samra Basharat ),( Ye Li ),( Zhonghu Bai ) 한국미생물 · 생명공학회 2021 Journal of microbiology and biotechnology Vol.31 No.11
Violacein, a purple pigment first isolated from a gram-negative coccobacillus Chromobacterium violaceum, has gained extensive research interest in recent years due to its huge potential in the pharmaceutic area and industry. In this review, we summarize the latest research advances concerning this pigment, which include (1) fundamental studies of its biosynthetic pathway, (2) production of violacein by native producers, apart from C. violaceum, (3) metabolic engineering for improved production in heterologous hosts such as Escherichia coli, Citrobacter freundii, Corynebacterium glutamicum, and Yarrowia lipolytica, (4) biological/pharmaceutical and industrial properties, (5) and applications in synthetic biology. Due to the intrinsic properties of violacein and the intermediates during its biosynthesis, the prospective research has huge potential to move this pigment into real clinical and industrial applications.
Rapid Screen of IL-5/IL-5Rα Blocking Antibodies in the HEK293-IL-5Rα-CSF2RB Transfected Cell Line
Shijie Li,Fei Han,Chang Liu,Weiyan Dai,Wenfeng Ke,Yongqi Chen,Eric Fordjour,Yankun Yang,Zhonghu Bai 한국생물공학회 2023 Biotechnology and Bioprocess Engineering Vol.28 No.4
Interleukin-5 (IL-5) binding to interleukin-5 receptor subunit alpha (IL-5Rα) increases the number of eosinophils and enhances eosinophil activity. This leads to eosinophil tissue infiltration and damage to the lungs, ultimately resulting in exacerbation of asthma. Antibodies that block IL-5 binding to IL-5Rα are thought to play an important role in advanced asthma. Currently, key methods used to screen for targeted drugs are Surface Plasmon Resonance which is costly and anti-proliferation assays which are tedious and have a low signal-to-noise ratio. Here we describe a Fluorescence Activated Cell Sorting (FACS) assay, based on human embryonic kidney (HEK)- 293 cells with stable expression of IL-5Rα and the cytokine receptor common subunit beta (CSF2RB). Cells co-expressing IL-5Rα and CSF2RB had a 16% increase in the ability to bind IL-5 compared to cells expressing only IL-5Rα. The optimal concentration of IL-5 for the FACS assay was 0.1 μg/mL. The established FACS was used to screen anti IL-5 nanobodies and hybridoma supernatants for candidate antibodies that block the IL-5/IL-5α interaction. When compared to anti-proliferation assays, this method saved up to 90% of the assay time, offering the advantage of rapidity and accuracy in vitro. The assay described here provides a novel approach for rapid screening of IL-5/IL- 5Rα blocking antibodies in vitro to accelerate the development of drugs for asthma.