Products from BPS Bioscience require a minimum order value above 400€Applications: Site-specific labeling of recombinant antibodies.
Background: Staphylococcal Sortase A is a bacterial transpeptidase that covalently attaches proteins to the bacterial cell wall, maintaining bacterial virulence and infectivity. Sortase A cleaves a specific peptide sequence (LPXTG recognition motif) within a target protein between threonine and glycine, with a strong preference for terminal locations in proteins. The cysteine residue of the active site forms a transient thioacyl intermediate complex with the substrate protein. This intermediate complex is then immediately attacked by oligo-glycine nucleophiles present on peptide-glycans of the bacterial wall to form an amide bond. This transpeptidase activity can be used for protein labeling. Using a recombinantly expressed antibody containing a C-terminal Sortase recognition sequence (LPXTG for S. aureus Sortase A) and a highly active Sortase A Heptamutant, the direct conjugation of poly-Glycine (G)
n-labels to the target protein can be achieved.
Description: The SiMPLe CHC Antibody Labeling Kit leverages Sortase Mediated Protein Ligation to specifically label recombinant antibodies containing the sequence LPXTG at the C-terminus of the heavy chain (CHC). A diverse number of poly-Glycine (G)
n molecules and labels (e.g. fluorophores, biotin, enzymes, peptides, etc.) are compatible with this kit. The site-specific conjugation ensures that the antigen binding site remains available for target binding and reduces heterogeneity compared to chemical conjugation methods, making it ideal for Antibody-Drug Conjugate (ADC) development and flow cytometry. The kit provides reagents to label 3 x 100 µg of recombinant antibody, as well as purification columns to aid in the removal of excess poly-Glycine (G)
n-label. Anti-HER2-LPETGH
6 and GGGK-FITC are included as positive controls.
Figure 1: Illustration of the mechanism of targeted labeling using the SiMPLe CHC Antibody Labeling Kit (Sortase Mediated Protein Ligation).
Each antibody contains two Sortase recognition sequences, so a maximum of two labels per antibody can be achieved
Storage Stability: This assay kit will perform optimally for up to 6 months from date of receipt when the materials are stored as directed
Warnings: Avoid freeze/thaw cycles
Biosafety Level: Not applicable (BSL-1)
References: Popp M., 2015 Methods Mol Biol. 1266:185-98.
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