Applications of novel affinity cassette methods: use of peptide fusion handles for the purification of recombinant proteins.
In this article, recent progress related to the use of different types of polypeptide fusion handles or 'tags' for the purification of recombinant proteins are critically discussed. In addition, novel aspects of the molecular cassette concept are elaborated, together with areas of potential application of these fundamental principles in molecular recognition. As evident from this review, the use of these concepts provides a powerful strategy for the high throughput isolation and purification of recombinant proteins and their derived domains, generated from functional genomic or zeomic studies, as part of the bioprocess technology leading to their commercial development, and in the study of molecular recognition phenomena per se. In addition, similar concepts can be exploited for high sensitivity analysis and detection, for the characterisation of protein bait/prey interactions at the molecular level, and for the immobilisation and directed orientation of proteins for use as biocatalysts/biosensors.
Hearn
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Centre for Bioprocess Technology, Department of Biochemistry and Molecular Biology, Monash University, Wellington Road, Clayton 3800 Australia. milton.hearn@med.monash.edu.au
Acosta
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D000345:Affinity Labels / Q000737:chemistry*; D000595:Amino Acid Sequence; D000915:Antibody Affinity; D001426:Bacterial Proteins / Q000235:genetics / Q000302:isolation & purification; D001435:Bacteriophages / Q000235:genetics; D001665:Binding Sites; D001710:Biotin; D002846:Chromatography, Affinity / Q000379:methods; D006639:Histidine / Q000235:genetics; D002479:Inclusion Bodies; D008320:Maltose / Q000378:metabolism; D008969:Molecular Sequence Data; D019151:Peptide Library; D015202:Protein Engineering / Q000379:methods; D017510:Protein Folding; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000737:chemistry* / Q000302:isolation & purification* / Q000378:metabolism; D013205:Staphylococcal Protein A / Q000235:genetics / Q000302:isolation & purification; D019809:Streptavidin
D000345:Affinity Labels; D001426:Bacterial Proteins; C034284:IgG Fc-binding protein, Streptococcus; D019151:Peptide Library; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D013205:Staphylococcal Protein A; D006639:Histidine; D008320:Maltose; D001710:Biotin; D019809:Streptavidin