Separating full-length protein from aggregating proteolytic products using filter flow-through purification.
Separation of full-length protein from proteolytic products is challenging, since the properties used to isolate the protein can also be present in proteolytic products. Many separation techniques risk non-specific protein adhesion and/or require a lot of time, enabling continued proteolysis and aggregation after lysis. We demonstrate that proteolytic products aggregate for two different proteins. As a result, full-length protein can be rapidly separated from these fragments by filter flow-through purification, resulting in a substantial protein purity enhancement. This rapid approach is likely to be useful for intrinsically disordered proteins, whose repetitive sequence composition and flexible nature can facilitate aggregation.
Department of Molecular and Cellular Medicine, Texas A&M Health Science Center, College Station, TX 77843-1114, USA.
Kelly A
KA
Churion
Department of Molecular and Cellular Medicine, Texas A&M Health Science Center, College Station, TX 77843-1114, USA.
Robert E
RE
Rogers
Department of Molecular and Cellular Medicine, Texas A&M Health Science Center, College Station, TX 77843-1114, USA.
Kayla J
KJ
Bayless
Department of Molecular and Cellular Medicine, Texas A&M Health Science Center, College Station, TX 77843-1114, USA. Electronic address: sebondos@tamhsc.edu.
Sarah E
SE
Bondos
D001671:Biochemistry / Q000295:instrumentation / Q000379:methods*; D002352:Carrier Proteins / Q000235:genetics / Q000302:isolation & purification; D029721:Drosophila Proteins / Q000235:genetics / Q000302:isolation & purification; D018398:Homeodomain Proteins / Q000235:genetics / Q000302:isolation & purification; D064267:Intrinsically Disordered Proteins / Q000302:isolation & purification; D011506:Proteins / Q000302:isolation & purification*; D014157:Transcription Factors / Q000235:genetics / Q000302:isolation & purification
D002352:Carrier Proteins; D029721:Drosophila Proteins; D018398:Homeodomain Proteins; D064267:Intrinsically Disordered Proteins; D011506:Proteins; C100642:SEC14L1 protein, human; D014157:Transcription Factors; C066177:Ubx protein, Drosophila
Aggregation; Filtration; Intrinsically disordered protein; Precipitation; Protein purification; Proteolysis