Simulation modeling of pooling for combinatorial protein engineering

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dc.contributor.authorPolizzi, KMko
dc.contributor.authorSpencer, CUko
dc.contributor.authorDubey, Ako
dc.contributor.authorMatsumura, Iko
dc.contributor.authorLee, JayHyungko
dc.contributor.authorRealff, MJko
dc.contributor.authorBommarius, ASko
dc.date.accessioned2013-03-08T10:35:46Z-
dc.date.available2013-03-08T10:35:46Z-
dc.date.created2012-02-06-
dc.date.created2012-02-06-
dc.date.issued2005-12-
dc.identifier.citationJOURNAL OF BIOMOLECULAR SCREENING, v.10, no.8, pp.856 - 864-
dc.identifier.issn1087-0571-
dc.identifier.urihttp://hdl.handle.net/10203/92848-
dc.description.abstractPooling in directed-evolution experiments will greatly increase the throughput of screening systems, but important parameters such as the number of good mutants created and the activity level increase of the good mutants will depend highly on the protein being engineered. The authors developed and validated a Monte Carlo simulation model of pooling that allows the testing of various scenarios in silico before starting experimentation. Using a simplified test system of 2 enzymes, beta-galactosidase (supermutant, or greatly improved enzyme) and beta-glucuronidase (dud, or enzyme with ancestral level of activity), the model accurately predicted the number of supermutants detected in experiments within a factor of 2. Additional simulations using more complex activity distributions show the versatility of the model. Pooling is most suited to cases such as the directed evolution of new function in a protein, where the background level of activity is minimized, making it easier to detect small increases in activity level. Pooling is most successful when a sensitive assay is employed. Using the model will increase the throughput of screening procedures for directed-evolution experiments and thus lead to speedier engineering of proteins.-
dc.languageEnglish-
dc.publisherSAGE PUBLICATIONS INC-
dc.subjectIN-VITRO EVOLUTION-
dc.subjectBETA-GLUCURONIDASE-
dc.subjectDIRECTED EVOLUTION-
dc.subjectRAPID EVOLUTION-
dc.subjectACTIVE-SITE-
dc.subjectGALACTOSIDASE-
dc.subjectSTRATEGY-
dc.subjectDESIGNS-
dc.subjectTOOL-
dc.titleSimulation modeling of pooling for combinatorial protein engineering-
dc.typeArticle-
dc.identifier.wosid000234407000011-
dc.identifier.scopusid2-s2.0-29144520637-
dc.type.rimsART-
dc.citation.volume10-
dc.citation.issue8-
dc.citation.beginningpage856-
dc.citation.endingpage864-
dc.citation.publicationnameJOURNAL OF BIOMOLECULAR SCREENING-
dc.identifier.doi10.1177/10871057105280134-
dc.contributor.localauthorLee, JayHyung-
dc.contributor.nonIdAuthorPolizzi, KM-
dc.contributor.nonIdAuthorSpencer, CU-
dc.contributor.nonIdAuthorDubey, A-
dc.contributor.nonIdAuthorMatsumura, I-
dc.contributor.nonIdAuthorRealff, MJ-
dc.contributor.nonIdAuthorBommarius, AS-
dc.type.journalArticleArticle-
dc.subject.keywordAuthordirected evolution-
dc.subject.keywordAuthorhigh-throughput screening-
dc.subject.keywordAuthorMonte Carlo simulation-
dc.subject.keywordAuthorprotein engineering-
dc.subject.keywordPlusIN-VITRO EVOLUTION-
dc.subject.keywordPlusBETA-GLUCURONIDASE-
dc.subject.keywordPlusDIRECTED EVOLUTION-
dc.subject.keywordPlusRAPID EVOLUTION-
dc.subject.keywordPlusACTIVE-SITE-
dc.subject.keywordPlusGALACTOSIDASE-
dc.subject.keywordPlusSTRATEGY-
dc.subject.keywordPlusDESIGNS-
dc.subject.keywordPlusTOOL-
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