Abstract: An acid-resistant peptide solubility tag (an “inclusion body tag”) is provided that is effective in producing peptides of interest in an insoluble form. Fusion peptide constructs comprising the inclusion body tag fused to a peptide of interest are provided. An acid cleavable peptide moiety separates the inclusion body tag from the peptide of interest so that acid hydrolysis can be used during subsequent processing steps to separate the tag from the desired peptide of interest. The present inclusion body tag's resistance to acid hydrolysis facilitates easier and cleaner separation of the peptide of interest after acid hydrolysis. Specifically, a ketosteroid isomerase-derived inclusion body tag is provided that has been engineered to be more resistant to acid hydrolysis.

Abstract: A method is provided to identify short peptide tags, referred to here as inclusion body tags (IBTs), useful for the generation of insoluble fusion peptides. A library of genetic constructs were prepared encoding fusion peptides comprising an inclusion body tag of 10-50 contiguous amino acids from a full-length insoluble protein operably linked to a peptide of interest. The library was designed to include a sufficient number of overlapping inclusion body tags to ensure that the entire length of the full-length insoluble protein was represented. Host cells transformed and expressing the genetic constructs were evaluated for inclusion body formation.

Abstract: Peptide tags, referred to here as inclusion body tags, are disclosed and are useful for the generation of insoluble fusion peptides. The fusion peptides comprise at least one inclusion body tag operably linked to a peptide of interest. Expression of the fusion peptide in a host cell results in a product that is insoluble and contained within inclusion bodies in the cell and/or cell lysate. The inclusion bodies may then be purified and the protein of interest may be isolated after cleavage from the inclusion body tag.

Abstract: Peptide tags, referred to here as inclusion body tags, are disclosed useful for the generation of insoluble fusion peptides. The fusion peptides comprise at least one inclusion body tag operably linked to a peptide of interest. Expression of the fusion peptide in a host cell results in a product that is insoluble and contained within inclusion bodies in the cell and/or cell lysate. The inclusion bodies may then be purified and the protein of interest may be isolated after cleavage from the inclusion body tag.

Abstract: Zein-based peptide tags, referred to here as inclusion body tags (IBTs), are disclosed useful for the generation of insoluble fusion peptides. The fusion peptides comprise at least one inclusion body tag operably linked to a peptide of interest. Expression of the fusion peptide in a host cell results in a product that is insoluble and contained within inclusion bodies in the cell and/or cell lysate. The inclusion bodies may then be purified and the protein of interest may be isolated after cleavage from the inclusion body tag.

Abstract: Peptide tags, referred to here as inclusion body tags, are disclosed useful for the generation of insoluble fusion peptides. The fusion peptides comprise at least one inclusion body tag operably linked to a peptide of interest. Expression of the fusion peptide in a host cell results in a product that is insoluble and contained within inclusion bodies in the cell and/or cell lysate. The inclusion bodies may then be purified and the protein of interest may be isolated after cleavage from the inclusion body tag.

Abstract: Peptide tags, referred to here as inclusion body tags, are disclosed useful for the generation of insoluble fusion peptides. The fusion peptides comprise at least one inclusion body tag operably linked to a peptide of interest. Expression of the fusion peptide in a host cell results in a product that is insoluble and contained within inclusion bodies in the cell and/or cell lysate. The inclusion bodies may then be purified and the protein of interest may be isolated after cleavage from the inclusion body tag.

Abstract: Cystatin-based peptide tags, referred to here as inclusion body tags (IBTs), are disclosed useful for the generation of insoluble fusion peptides. The fusion peptides comprise at least one inclusion body tag operably linked to a peptide of interest. Expression of the fusion peptide in a host cell results in a product that is insoluble and contained within inclusion bodies in the cell and/or cell lysate. The inclusion bodies may then be purified and the protein of interest may be isolated after cleavage from the inclusion body tag.

Abstract: Peptide tags, referred to here as inclusion body tags, are disclosed and are useful for the generation of insoluble fusion peptides. The fusion peptides comprise at least one inclusion body tag operably linked to a peptide of interest. Expression of the fusion peptide in a host cell results in a product that is insoluble and contained within inclusion bodies in the cell and/or cell lysate. The inclusion bodies may then be purified and the protein of interest may be isolated after cleavage from the inclusion body tag.

Abstract: The invention relates to the recombinant expression of a peptide of interest in the form of a fusion protein comprising a solubility tag. The fusion protein comprises at least two portions separated by a cleavable peptide sequence wherein one portion is devoid of cysteine residues and the second portion comprises an effective number of cross-linkable cysteine residues. After cell lysis and isolation of the fusion protein, the fusion protein is subsequently cleaved into a mixture of first and second portions. Oxidative cross-linking is used to selectively precipitate one of the two portions to facilitate simple and effective separation of the peptide of interest.

Abstract: An acid-resistant peptide solubility tag (an “inclusion body tag”) is provided that is effective in producing peptides of interest in an insoluble form. Fusion peptide constructs comprising the inclusion body tag fused to a peptide of interest are provided. An acid cleavable peptide moiety separates the inclusion body tag from the peptide of interest so that acid hydrolysis can be used during subsequent processing steps to separate the tag from the desired peptide of interest. The present inclusion body tag's resistance to acid hydrolysis facilitates easier and cleaner separation of the peptide of interest after acid hydrolysis. Specifically, a ketosteroid isomerase-derived inclusion body tag is provided that has been engineered to be more resistant to acid hydrolysis.

Abstract: Peptide tags, referred to here as inclusion body tags, are disclosed useful for the generation of insoluble fusion peptides. The fusion peptides comprise at least one inclusion body tag operably linked to a peptide of interest. Expression of the fusion peptide in a host cell results in a product that is insoluble and contained within inclusion bodies in the cell and/or cell lysate. The inclusion bodies may then be purified and the protein of interest may be isolated after cleavage from the inclusion body tag.

Abstract: Peptide tags, referred to here as inclusion body tags, are disclosed and are useful for the generation of insoluble fusion peptides. The fusion peptides comprise at least one inclusion body tag operably linked to a peptide of interest. Expression of the fusion peptide in a host cell results in a product that is insoluble and contained within inclusion bodies in the cell and/or cell lysate. The inclusion bodies may then be purified and the protein of interest may be isolated after cleavage from the inclusion body tag.

Abstract: Peptide tags, referred to here as inclusion body tags, are disclosed and are useful for the generation of insoluble fusion peptides. The fusion peptides comprise at least one inclusion body tag operably linked to a peptide of interest. Expression of the fusion peptide in a host cell results in a product that is insoluble and contained within inclusion bodies in the cell and/or cell lysate. The inclusion bodies may then be purified and the protein of interest may be isolated after cleavage from the inclusion body tag.

Abstract: Zein-based peptide tags, referred to here as inclusion body tags (IBTs), are disclosed useful for the generation of insoluble fusion peptides. The fusion peptides comprise at least one inclusion body tag operably linked to a peptide of interest. Expression of the fusion peptide in a host cell results in a product that is insoluble and contained within inclusion bodies in the cell and/or cell lysate. The inclusion bodies may then be purified and the protein of interest may be isolated after cleavage from the inclusion body tag.

Abstract: Cystatin-based peptide tags, referred to here as inclusion body tags (IBTs), are disclosed useful for the generation of insoluble fusion peptides. The fusion peptides comprise at least one inclusion body tag operably linked to a peptide of interest. Expression of the fusion peptide in a host cell results in a product that is insoluble and contained within inclusion bodies in the cell and/or cell lysate. The inclusion bodies may then be purified and the protein of interest may be isolated after cleavage from the inclusion body tag.