当前位置：
X-MOL 学术
›
Biochemistry
›
论文详情
Our official English website, www.x-mol.net, welcomes your
feedback! (Note: you will need to create a separate account there.)
Characterization of an Antibody Recognizing the Conserved Inner Core of Pseudomonas aeruginosa Lipopolysaccharides
Biochemistry ( IF 2.9 ) Pub Date : 2020-10-21 , DOI: 10.1021/acs.biochem.0c00642 Stefano Elli 1 , Anna Alekseeva 2 , Boopathy Ramakrishnan 3 , Tyree Koch 3 , Andrew Wollacott 3 , Karthik Viswanathan 3 , Kai Li 3 , James C. Delaney 3 , Zachary Shriver 3 , Obadiah Plante 3 , Marco Guerrini 1
Biochemistry ( IF 2.9 ) Pub Date : 2020-10-21 , DOI: 10.1021/acs.biochem.0c00642 Stefano Elli 1 , Anna Alekseeva 2 , Boopathy Ramakrishnan 3 , Tyree Koch 3 , Andrew Wollacott 3 , Karthik Viswanathan 3 , Kai Li 3 , James C. Delaney 3 , Zachary Shriver 3 , Obadiah Plante 3 , Marco Guerrini 1
Affiliation
|
Bacterial infections are a growing public health threat with carbapenem-resistant Pseudomonas aeruginosa being classified as a Priority 1 critical threat by the World Health Organization. Antibody-based therapeutics can serve as an alternative and in some cases supplement antibiotics for the treatment of bacterial infections. The glycans covering the bacterial cell surface have been proposed as intriguing targets for binding by antibodies; however, antibodies that can engage with high affinity and specificity with glycans are much less common compared to antibodies that engage with protein antigens. In this study, we sought to characterize an antibody that targets a conserved glycan epitope on the surface of Pseudomonas. First, we characterized the breadth of binding of VSX, demonstrating that the VSX is specific to Pseudomonas but can bind across multiple serotypes of the organism. Next, we provide insight into how VSX engages with its target epitope, using a combination of biolayer interferometry and nuclear magnetic resonance, and verify our results using site-directed mutagenesis experiments. We demonstrate that the antibody, with limited somatic hypermutation of the complementarity-determining regions (CDRs) and with a characteristic set of arginines within the CDRs, specifically targets the conserved inner core of Pseudomonas lipopolysaccharides. Our results provide important additional context to antibody–glycan contacts and provide insight useful for the construction of vaccines and therapeutics against Pseudomonas aeruginosa, an important human pathogen.
中文翻译：
识别铜绿假单胞菌脂多糖的保守内核的抗体的表征
细菌感染是一种日益严重的公共卫生威胁，耐碳青霉烯的铜绿假单胞菌被世界卫生组织列为优先重点之一。基于抗体的疗法可以替代，在某些情况下可以补充抗生素来治疗细菌感染。已经提出了覆盖细菌细胞表面的聚糖作为吸引人的抗体结合靶标。然而，与聚糖具有高亲和力和特异性的抗体相比，与蛋白质抗原相结合的抗体要少得多。在这项研究中，我们试图表征一种针对假单胞菌表面上保守的聚糖表位的抗体。首先，我们表征了VSX结合的广度，表明VSX对假单胞菌具有特异性，但可以跨生物的多种血清型结合。接下来，我们结合生物层干涉测量法和核磁共振技术，深入了解VSX如何与其目标表位结合，并使用定点诱变实验验证我们的结果。我们证明，该抗体，具有有限的体细胞互补决定区（CDR）的超突变，并在CDR中具有精氨酸的特征集，专门针对假单胞菌的保守内核脂多糖。我们的结果为抗体-聚糖的接触提供了重要的额外环境，并为构建抗铜绿假单胞菌（一种重要的人类病原体）的疫苗和治疗方法提供了有用的见识。
更新日期：2020-11-03
中文翻译：
识别铜绿假单胞菌脂多糖的保守内核的抗体的表征
细菌感染是一种日益严重的公共卫生威胁，耐碳青霉烯的铜绿假单胞菌被世界卫生组织列为优先重点之一。基于抗体的疗法可以替代，在某些情况下可以补充抗生素来治疗细菌感染。已经提出了覆盖细菌细胞表面的聚糖作为吸引人的抗体结合靶标。然而，与聚糖具有高亲和力和特异性的抗体相比，与蛋白质抗原相结合的抗体要少得多。在这项研究中，我们试图表征一种针对假单胞菌表面上保守的聚糖表位的抗体。首先，我们表征了VSX结合的广度，表明VSX对假单胞菌具有特异性，但可以跨生物的多种血清型结合。接下来，我们结合生物层干涉测量法和核磁共振技术，深入了解VSX如何与其目标表位结合，并使用定点诱变实验验证我们的结果。我们证明，该抗体，具有有限的体细胞互补决定区（CDR）的超突变，并在CDR中具有精氨酸的特征集，专门针对假单胞菌的保守内核脂多糖。我们的结果为抗体-聚糖的接触提供了重要的额外环境，并为构建抗铜绿假单胞菌（一种重要的人类病原体）的疫苗和治疗方法提供了有用的见识。
京公网安备 11010802027423号