Matches in SemOpenAlex for { <https://semopenalex.org/work/W2143867882> ?p ?o ?g. }
- W2143867882 endingPage "232" @default.
- W2143867882 startingPage "221" @default.
- W2143867882 abstract "Integration of the chemical extraction of recombinant inclusion-body protein from Escherichia coli, and its recovery by metal-affinity expanded-bed adsorption (IMAC-EBA) under denaturing conditions, was investigated. The viral coat protein L1 with a hexa-histidine tag was expressed in Escherichia coli HMS174(DE3) as a model protein. Interference of released host DNA with adsorbent fluidization in the EBA step was solved by selective precipitation using spermine and low-speed centrifugation. However, the capacity and selectivity of the adsorbent for L1 remained lower than anticipated. The binding of L1 to immobilized Ni(2+) was therefore studied in detail using surface plasmon resonance (SPR). The Tris buffer and ethylene-diamine tetraacetic acid (EDTA) used in the extraction mixture were found to interfere significantly with the L1-Ni(2+) interaction. The SPR studies suggest that L1 binding could be improved by replacing the Tris buffer with HEPES and by adding CaCl(2) to inactivate the EDTA. The modified chemical extraction conditions resulted in effective L1 extraction from cytoplasmic inclusion bodies, at high cell density (OD(600 )= 80) and without the use of reducing agent, into a medium optimized for subsequent IMAC recovery. The modified buffer conditions resulted in an improved binding capacity and a good L1 purification factor (12.7) and recovery yield (71%). This work demonstrates that it is possible to reduce the complexity and hence the cost associated with traditional processes used to prepare purified denatured protein, ready for refolding, from cytoplasmic inclusion bodies." @default.
- W2143867882 created "2016-06-24" @default.
- W2143867882 creator A5021586160 @default.
- W2143867882 creator A5030698014 @default.
- W2143867882 creator A5044210377 @default.
- W2143867882 creator A5048165624 @default.
- W2143867882 date "2002-11-25" @default.
- W2143867882 modified "2023-09-23" @default.
- W2143867882 title "Coupling of chemical extraction and expanded-bed adsorption for simplified inclusion-body processing: Optimization using surface plasmon resonance" @default.
- W2143867882 cites W1508622807 @default.
- W2143867882 cites W1846731397 @default.
- W2143867882 cites W1978519233 @default.
- W2143867882 cites W1987350137 @default.
- W2143867882 cites W1990429347 @default.
- W2143867882 cites W2003358293 @default.
- W2143867882 cites W2003470884 @default.
- W2143867882 cites W2038315892 @default.
- W2143867882 cites W2044543494 @default.
- W2143867882 cites W2069118589 @default.
- W2143867882 cites W2095862282 @default.
- W2143867882 cites W2107391580 @default.
- W2143867882 cites W2111668132 @default.
- W2143867882 cites W2115440372 @default.
- W2143867882 cites W2119471163 @default.
- W2143867882 cites W2120628792 @default.
- W2143867882 cites W2121819501 @default.
- W2143867882 cites W2123631149 @default.
- W2143867882 cites W2138796104 @default.
- W2143867882 cites W2145892424 @default.
- W2143867882 cites W2162361391 @default.
- W2143867882 cites W4238854347 @default.
- W2143867882 cites W4246745169 @default.
- W2143867882 cites W4251401925 @default.
- W2143867882 cites W4377854697 @default.
- W2143867882 doi "https://doi.org/10.1002/bit.10471" @default.
- W2143867882 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/12451558" @default.
- W2143867882 hasPublicationYear "2002" @default.
- W2143867882 type Work @default.
- W2143867882 sameAs 2143867882 @default.
- W2143867882 citedByCount "24" @default.
- W2143867882 countsByYear W21438678822012 @default.
- W2143867882 countsByYear W21438678822013 @default.
- W2143867882 countsByYear W21438678822018 @default.
- W2143867882 crossrefType "journal-article" @default.
- W2143867882 hasAuthorship W2143867882A5021586160 @default.
- W2143867882 hasAuthorship W2143867882A5030698014 @default.
- W2143867882 hasAuthorship W2143867882A5044210377 @default.
- W2143867882 hasAuthorship W2143867882A5048165624 @default.
- W2143867882 hasConcept C104317684 @default.
- W2143867882 hasConcept C106847996 @default.
- W2143867882 hasConcept C150394285 @default.
- W2143867882 hasConcept C155672457 @default.
- W2143867882 hasConcept C171250308 @default.
- W2143867882 hasConcept C178790620 @default.
- W2143867882 hasConcept C185592680 @default.
- W2143867882 hasConcept C192562407 @default.
- W2143867882 hasConcept C207891110 @default.
- W2143867882 hasConcept C2781014771 @default.
- W2143867882 hasConcept C43617362 @default.
- W2143867882 hasConcept C4725764 @default.
- W2143867882 hasConcept C547475151 @default.
- W2143867882 hasConcept C55493867 @default.
- W2143867882 hasConcept C88001094 @default.
- W2143867882 hasConcept C8985478 @default.
- W2143867882 hasConceptScore W2143867882C104317684 @default.
- W2143867882 hasConceptScore W2143867882C106847996 @default.
- W2143867882 hasConceptScore W2143867882C150394285 @default.
- W2143867882 hasConceptScore W2143867882C155672457 @default.
- W2143867882 hasConceptScore W2143867882C171250308 @default.
- W2143867882 hasConceptScore W2143867882C178790620 @default.
- W2143867882 hasConceptScore W2143867882C185592680 @default.
- W2143867882 hasConceptScore W2143867882C192562407 @default.
- W2143867882 hasConceptScore W2143867882C207891110 @default.
- W2143867882 hasConceptScore W2143867882C2781014771 @default.
- W2143867882 hasConceptScore W2143867882C43617362 @default.
- W2143867882 hasConceptScore W2143867882C4725764 @default.
- W2143867882 hasConceptScore W2143867882C547475151 @default.
- W2143867882 hasConceptScore W2143867882C55493867 @default.
- W2143867882 hasConceptScore W2143867882C88001094 @default.
- W2143867882 hasConceptScore W2143867882C8985478 @default.
- W2143867882 hasIssue "2" @default.
- W2143867882 hasLocation W21438678821 @default.
- W2143867882 hasLocation W21438678822 @default.
- W2143867882 hasOpenAccess W2143867882 @default.
- W2143867882 hasPrimaryLocation W21438678821 @default.
- W2143867882 hasRelatedWork W1972750290 @default.
- W2143867882 hasRelatedWork W2060806242 @default.
- W2143867882 hasRelatedWork W2093393366 @default.
- W2143867882 hasRelatedWork W2123740930 @default.
- W2143867882 hasRelatedWork W2143867882 @default.
- W2143867882 hasRelatedWork W2354563564 @default.
- W2143867882 hasRelatedWork W2361386354 @default.
- W2143867882 hasRelatedWork W2396027010 @default.
- W2143867882 hasRelatedWork W2949375571 @default.
- W2143867882 hasRelatedWork W2951877688 @default.
- W2143867882 hasVolume "81" @default.
- W2143867882 isParatext "false" @default.
- W2143867882 isRetracted "false" @default.