B. Bheemareddy, P. Iyer, Kranthi Vemparala, V. R. Dirisala
{"title":"A simple and novel sample preparation approach for effective characterization of antibody low molecular weight impurities by CE-SDS method","authors":"B. Bheemareddy, P. Iyer, Kranthi Vemparala, V. R. Dirisala","doi":"10.5530/CTBP.2020.3.32","DOIUrl":null,"url":null,"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.","PeriodicalId":10980,"journal":{"name":"Current Trends in Biotechnology and Pharmacy","volume":"14 1","pages":"311-318"},"PeriodicalIF":0.0000,"publicationDate":"2020-09-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Current Trends in Biotechnology and Pharmacy","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.5530/CTBP.2020.3.32","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"Pharmacology, Toxicology and Pharmaceutics","Score":null,"Total":0}
引用次数: 0
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.
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.