Pete Gagnon, Chia-Wei Cheung, Eric J Lepin, Anna M Wu, Mark A Sherman, Andrew A Raubitschek, Paul J Yazaki
{"title":"Minibodies and Multimodal Chromatography Methods: A Convergence of Challenge and Opportunity.","authors":"Pete Gagnon, Chia-Wei Cheung, Eric J Lepin, Anna M Wu, Mark A Sherman, Andrew A Raubitschek, Paul J Yazaki","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>This case study describes early phase purification process development for a recombinant anticancer minibody produced in mammalian cell culture. The minibody did not bind to protein A. Cation-exchange, anion-exchange, hydrophobic-interaction, and hydroxyapatite (eluted by phosphate gradient) chromatographic methods were scouted, but the minibody coeluted with BSA to a substantial degree on each. Hydroxyapatite eluted with a sodium chloride gradient separated BSA and also removed a dimeric contaminant, but BSA consumed so much binding capacity that this proved impractical as a capture tool. Capto MMC media proved capable of supporting adequate capture and significant dimer removal, although both loading and elution selectivity varied dramatically with the amount of supernatant applied to the column. An anion-exchange step was included to fortify overall virus and DNA removal. These results illustrate the value of multimodal chromatography methods when affinity chromatography methods are lacking and conventional alternatives prove inadequate.</p>","PeriodicalId":89332,"journal":{"name":"BioProcess international","volume":"8 2","pages":"26-35"},"PeriodicalIF":0.0,"publicationDate":"2010-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3187938/pdf/nihms319083.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"30195679","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Mary S Rosendahl, Daniel H Doherty, Darin J Smith, Alison M Bendele, George N Cox
{"title":"Site-Specific Protein PEGylation: Application to Cysteine Analogs of Recombinant Human Granulocyte Colony-Stimulating Factor.","authors":"Mary S Rosendahl, Daniel H Doherty, Darin J Smith, Alison M Bendele, George N Cox","doi":"","DOIUrl":"","url":null,"abstract":"","PeriodicalId":89332,"journal":{"name":"BioProcess international","volume":"3 4","pages":"52-60"},"PeriodicalIF":0.0,"publicationDate":"2005-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5995563/pdf/nihms907980.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"36221233","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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