کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
5916720 | 1163753 | 2015 | 9 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Effect of protein aggregates on characterization of FcRn binding of Fc-fusion therapeutics
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کلمات کلیدی
PBSAlphaScreenFcRnHMWSECFluorescence resonance energy transfer - انتقال انرژی رزونانس FluorescenceFRET - انتقال انرژی رزونانسی فورسترBiolayer Interferometry - اینترفا متر BiilayerELISA - تست الیزاEnzyme-linked immunosorbent assay - تست الیزاSurface plasmon resonance - تشدید پلاسمون سطحیSPR - تشدید پلاسمون سطحیAssociation constant - ثابت انجمنDissociation constant - حد تفکیکBLI - داشته باشد،Phosphate buffered saline - فسفات بافر شورMeS - مسAggregates - مصالحultraviolet light - نور ماوراءبنفش یا نور فرابنفشSize exclusion chromatography - کروماتوگرافی اندازهای طردیhigh-pressure liquid chromatography - کروماتوگرافی مایع با فشار بالاHPLC - کروماتوگرافی مایعی کاراneonatal Fc receptor - گیرنده Fc نوزادان
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
زیست شناسی مولکولی
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چکیده انگلیسی
Recycling of antibodies and Fc containing therapeutic proteins by the neonatal Fc receptor (FcRn) is known to prolong their persistence in the bloodstream. Fusion of Fc fragment of IgG1 to other proteins is one of the strategies to improve their pharmacokinetic properties. Accurate measurement of Fc-FcRn binding provides information about the strength of this interaction, which in most cases correlates with serum half-life of the protein. It can also offer insight into functional integrity of Fc region. We investigated FcRn binding activity of a large set of Fc-fusion samples after thermal stress by the method based on AlphaScreen technology. An unexpected significant increase in FcR binding was found to correlate with formation of aggregates in these samples. Monomer purified from a thermally-stressed sample had normal FcRn binding, confirming that its Fc portion was intact. Experiments with aggregates spiked into a sample with low initial aggregation level, demonstrated strong correlation between the level of aggregates and FcRn binding. This correlation varied significantly in different methods. By introducing modifications to the assay format we were able to minimize the effects of aggregated species on FcRn binding, which should prevent masking functional changes of Fc-fusion protein. Biolayer interferometry (BLI) was used as an alternative method to measure FcRn binding. Both optimized AlphaScreen- and BLI-based assays were sensitive to structural changes in Fc portion of the molecule, such as oxidation of methionines 252 and 428, and therefore suitable for characterization of FcRn binding.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Molecular Immunology - Volume 67, Issue 2, Part B, October 2015, Pages 616-624
Journal: Molecular Immunology - Volume 67, Issue 2, Part B, October 2015, Pages 616-624
نویسندگان
Adriana Bajardi-Taccioli, Andrew Blum, Chongfeng Xu, Zoran Sosic, Svetlana Bergelson, Marina Feschenko,