A simple and novel sample preparation approach for effective characterization of antibody low molecular weight impurities by CE-SDS method

Q3 Pharmacology, Toxicology and Pharmaceutics Current Trends in Biotechnology and Pharmacy Pub Date : 2020-09-22 DOI:10.5530/CTBP.2020.3.32
B. Bheemareddy, P. Iyer, Kranthi Vemparala, V. R. Dirisala
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Abstract

Effect of sample buffer composition and pH on LMW impurities analysis Abstract Product and process related impurities of biopharmaceuticals have serious implications on product safety and efficacy in clinical use. Low molecular weight (LMW) impurities are generated during process and stability studies are routinely analyzed using non-reducing capillary electrophoresis with SDS during different stages of product development and release. The current sample processing methodology with heat denaturation is known to induce fragmentation and interfere with the LMW impurity analysis. In this study, we compared different sample processing buffers with different compositions and pH and finally found a solution to the problem of sample artifacts generated during heat denaturation step of sample processing which interferes with the LMW impurity analysis by CE-SDS method. We compared three sample buffers (100 mM Tris-Cl, 25mM Citrate and 25mM citrate with Urea) in for their ability to maintain product integrity during sample processing at different pH and temperatures in the non-reducing CE-SDS analysis. This study suggests that, the sample processing with 25mM citrate with 8M Urea sample processing buffer does not require heat denaturation at higher temperatures and hence is the most appropriate buffer for sample processing in the LMW impurity analysis. The 25mM Citrate + 8M Urea buffer has shown better drug product stability and integrity compared to other buffers. Hence, we recommend the 25mM Citrate buffer with 8M Urea for sample processing in LMW impurity analysis by CE-SDS method.
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CE-SDS法是一种简单、新颖的样品制备方法,可有效地表征抗体低分子量杂质
样品缓冲液组成和pH对LMW杂质分析的影响摘要生物制药的产品和工艺相关杂质对临床使用中的产品安全性和有效性有严重影响。低分子量(LMW)杂质在工艺过程中产生,在产品开发和释放的不同阶段,使用SDS的非还原毛细管电泳对稳定性研究进行常规分析。目前已知具有热变性的样品处理方法会导致碎片化并干扰LMW杂质分析。在本研究中,我们比较了不同组成和pH的不同样品处理缓冲液,最终找到了解决样品处理热变性步骤中产生的样品伪影问题的方法,该问题干扰了CE-SDS方法对LMW杂质的分析。在非还原性CE-SDS分析中,我们比较了三种样品缓冲液(100mM Tris-Cl、25mM柠檬酸盐和25mM柠檬酸酯与尿素)在不同pH和温度的样品处理过程中保持产品完整性的能力。本研究表明,用25mM柠檬酸盐和8M尿素样品处理缓冲液处理样品不需要在较高温度下进行热变性,因此是LMW杂质分析中最适合样品处理的缓冲液。与其他缓冲液相比,25mM柠檬酸盐+8M尿素缓冲液显示出更好的药品稳定性和完整性。因此,我们推荐使用含有8M尿素的25mM柠檬酸盐缓冲液,用于CE-SDS法LMW杂质分析的样品处理。
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Current Trends in Biotechnology and Pharmacy
Current Trends in Biotechnology and Pharmacy Pharmacology, Toxicology and Pharmaceutics-Pharmaceutical Science
CiteScore
0.70
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期刊介绍: The Association of Biotechnology and Pharmacy (ABAP) will be useful to form a forum for scientists so that they can bring together to discuss and find scientific solutions to the problems of society. The annual meetings will help the members to share their knowledge and publish their research knowledge particularly by members and fellows of the Association and special care will be taken to provide an opportunity for young scientists. Besides this the association is planned to organize symposia, seminars and workshops on current developments of Biotechnology and Pharmacy particularly on the subject of current scientific interest, and the proceedings of which will be published regularly. And in view of the vast development of science and to disseminate the problems in publication of research work, an international journal of Current Trends in Biotechnology and Pharmacy has been started by ABAP.
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