دورية أكاديمية
Characterization of the complex formed between a potent neutralizing ovine-derived polyclonal anti-TNFα Fab fragment and human TNFα.
| العنوان: | Characterization of the complex formed between a potent neutralizing ovine-derived polyclonal anti-TNFα Fab fragment and human TNFα. |
|---|---|
| المؤلفون: | Abbott WM; AstraZeneca, Alderley Park, Macclesfield, Cheshire SK10 4TG, U.K. mark.abbott@astrazeneca.com, Snow M, Eckersley S, Renshaw J, Davies G, Norman RA, Ceuppens P, Slootstra J, Benschop JJ, Hamuro Y, Lee JE, Newham P |
| المصدر: | Bioscience reports [Biosci Rep] 2013 Aug 23; Vol. 33 (4). Date of Electronic Publication: 2013 Aug 23. |
| نوع المنشور: | Journal Article; Research Support, Non-U.S. Gov't |
| اللغة: | English |
| بيانات الدورية: | Publisher: Portland Press on behalf of the Biochemical Society Country of Publication: England NLM ID: 8102797 Publication Model: Electronic Cited Medium: Internet ISSN: 1573-4935 (Electronic) Linking ISSN: 01448463 NLM ISO Abbreviation: Biosci Rep Subsets: MEDLINE |
| أسماء مطبوعة: | Publication: London : Portland Press on behalf of the Biochemical Society Original Publication: London : The Biochemical Society, c1981- |
| مواضيع طبية MeSH: | Immunoglobulin Fab Fragments/*chemistry , Tumor Necrosis Factor-alpha/*chemistry, Amino Acid Sequence ; Amino Acid Substitution ; Animals ; Cell Line ; Cell Survival/drug effects ; Epitope Mapping ; Epitopes/chemistry ; Epitopes/immunology ; Humans ; Immunoglobulin Fab Fragments/pharmacology ; Mice ; Molecular Sequence Data ; Molecular Weight ; Mutagenesis, Site-Directed ; Protein Binding ; Sheep, Domestic ; Tumor Necrosis Factor-alpha/antagonists & inhibitors ; Tumor Necrosis Factor-alpha/genetics ; Tumor Necrosis Factor-alpha/immunology |
| مستخلص: | TNFα (tumour necrosis factor α) is an early mediator in the systemic inflammatory response to infection and is therefore a therapeutic target in sepsis. AZD9773 is an ovine-derived, polyclonal anti-TNFα Fab fragment derived from a pool of serum and currently being developed as a treatment for severe sepsis and septic shock. In the present study, we show that although AZD9773 has a modest affinity for TNFα in a binding assay, the Ki in a cell-based assay is approximately four orders of magnitude lower. We show using SEC (size exclusion chromatography) that the maximum size of the complex between AZD9773 and TNFα is consistent with approximately 12 Fabs binding to one TNFα trimer. A number of approaches were taken to map the epitopes recognized by AZD9773. These revealed that a number of different regions on TNFα are involved in binding to the polyclonal Fab. The data suggest that there are probably three epitopes per monomer that are responsible for most of the inhibition by AZD9773 and that all three can be occupied at the same time in the complex. We conclude that AZD9773 is clearly demonstrated to bind to multiple epitopes on TNFα and suggest that the polyclonal nature may account, at least in part, for the very high potency observed in cell-based assays. |
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| المشرفين على المادة: | 0 (AZD9773) 0 (Epitopes) 0 (Immunoglobulin Fab Fragments) 0 (TNF protein, human) 0 (Tumor Necrosis Factor-alpha) |
| تواريخ الأحداث: | Date Created: 20130719 Date Completed: 20140421 Latest Revision: 20211021 |
| رمز التحديث: | 20231215 |
| مُعرف محوري في PubMed: | PMC3755337 |
| DOI: | 10.1042/BSR20130044 |
| PMID: | 23863106 |
| قاعدة البيانات: | MEDLINE |
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