The following SOPs detail expression and purification of the SARS-CoV-2 diagnostic assay reagents developed by UCAM as part of the AfriDx project. See also the details of WP3 and our training resources for more information.
SOPs for Engineered Boon-RTX1) Protein Expression of Engineered Boon-RTX2) Silica Purification of Engineered Boon-RTXBoon-RTX is a recombinantly expressed, engineered reverse transcriptase that catalyses RNA to DNA conversion. It has been fused with a silaffin-tag (R5) and mCherry reporter. This combination of proteins and tags allows for low-cost purification using silica and visual tracking through the prodcution process. The protein also has a His-Tag to provide a secondary purification option. |
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SOPs for Engineered Boon2-Bst1) Protein Expression of Engineered Boon2-Bst2) Autoinduction Expression of Engineered Boon2-Bst3) Silica Purification of Engineered Boon2-BstBoon-Bst is a recombinantly expressed, engineered strand displacing DNA polymerase that can be applied in LAMP-based DNA amplification. It has been fused with a silaffin-tag (R5) and mCherry reporter fusion. This combination of proteins and tags allows for low-cost purification using silica and visual tracking through the prodcution process. The protein also has a His-Tag to provide a secondary purification option. |
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SOPs for Engineered Boon-scFv1) Autoinduction Protein Expression of Boon-scFv Engineered Fluorescent Single Chained Antibody2) Purification of Boon-scFv Engineered Fluorescent Single Chained AntibodyBoon-scFv is a recombinantly expressed single-chain variable fragment (scFv) targeting human Immunoglobulin A/G/M for detecting immune responses to a SARS-CoV-2 infection. It has been synthesised with an internal mCherry for protein tracking, a pectate lyase B (pelB) leader sequence for improved productivity, a His-tag for purification, and a FLAG-tag for specific identification. |
