کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
10962922 | 1102657 | 2015 | 6 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
HPAEC-PAD quantification of Haemophilus influenzae type b polysaccharide in upstream and downstream samples
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کلمات کلیدی
PBSd-glucose-6-phosphateNaAcWFIglc-6-PRIVMUSPESTDPrPIPCHPAEC-PADkDaMWCODSPGlcHPSECNaOHcpswater for injection - آب برای تزریقSodium acetate - استات سدیمExternal standard - استاندارد خارجیNucleic acids - اسید نوکلئیکRib - دندهRetention time - زمان بازداریWorld Health Organization - سازمان بهداشت جهانیRefractive index - ضریب شکستUpstream processing - فرآیند بالادستDownstream processing - فرایند فرآیند پایینPhosphate buffered saline - فسفات بافر شورHaemophilus influenzae type b - نوع hemophilus influenzae bHib - هیبsodium hydroxide - هیدروکسید سدیمCapsular polysaccharide - پلی ساکارید کپسولPolysaccharide - چندقندی ها یا پلیساکاریدهاoptical density - چگالی نوریmolecular weight cut-off - کاهش وزن مولکولیanion exchange chromatography - کروماتوگرافی تبادل آنیونhigh performance size exclusion chromatography - کروماتوگرافی خروج با اندازه کافی بالاHigh pressure liquid chromatography - کروماتوگرافی مایع با فشار بالاhigh performance anion exchange chromatography with pulsed amperometric detection - کروماتوگرافی مبدل آنیون با عملکرد بالا با تشخیص آمپومتریک پالسWHO - کهKilo Dalton - کیلو دالتونGlucose - گلوکز
موضوعات مرتبط
علوم زیستی و بیوفناوری
ایمنی شناسی و میکروب شناسی
ایمونولوژی
پیش نمایش صفحه اول مقاله
چکیده انگلیسی
Due to the rapidly increasing introduction of Haemophilus influenzae type b (Hib) and other conjugate vaccines worldwide during the last decade, reliable and robust analytical methods are needed for the quantitative monitoring of intermediate samples generated during fermentation (upstream processing, USP) and purification (downstream processing, DSP) of polysaccharide vaccine components. This study describes the quantitative characterization of in-process control (IPC) samples generated during the fermentation and purification of the capsular polysaccharide (CPS), polyribosyl-ribitol-phosphate (PRP), derived from Hib. Reliable quantitative methods are necessary for all stages of production; otherwise accurate process monitoring and validation is not possible. Prior to the availability of high performance anion exchange chromatography methods, this polysaccharide was predominantly quantified either with immunochemical methods, or with the colorimetric orcinol method, which shows interference from fermentation medium components and reagents used during purification. Next to an improved high performance anion exchange chromatography-pulsed amperometric detection (HPAEC-PAD) method, using a modified gradient elution, both the orcinol assay and high performance size exclusion chromatography (HPSEC) analyses were evaluated. For DSP samples, it was found that the correlation between the results obtained by HPAEC-PAD specific quantification of the PRP monomeric repeat unit released by alkaline hydrolysis, and those from the orcinol method was high (R2Â =Â 0.8762), and that it was lower between HPAEC-PAD and HPSEC results. Additionally, HPSEC analysis of USP samples yielded surprisingly comparable results to those obtained by HPAEC-PAD. In the early part of the fermentation, medium components interfered with the different types of analysis, but quantitative HPSEC data could still be obtained, although lacking the specificity of the HPAEC-PAD method. Thus, the HPAEC-PAD method has the advantage of giving a specific response compared to the orcinol assay and HPSEC, and does not show interference from various components that can be present in intermediate and purified PRP samples.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Vaccine - Volume 33, Issue 48, 27 November 2015, Pages 6908-6913
Journal: Vaccine - Volume 33, Issue 48, 27 November 2015, Pages 6908-6913
نویسندگان
Robert M.F. van der Put, Alex de Haan, Jan G.M. van den IJssel, Ahd Hamidi, Michel Beurret,