The expression and purification of the extracellular domain of human PSMA (residues 44–750) that comprised an N-terminal Avi-tag (Avi-PSMA) run according to protocol published earlier [54 (link),84 (link)]. Briefly, the recombinant protein was expressed by stably transfected Schneider S2 cells and purified by the combination of affinity chromatography (Streptavidin Mutein Matrix, Roche) and size-exclusion chromatography on a Superdex 200 column (GE Healthcare Bio-Sciences). Concentrated PSMA stock was stored at −80 °C until further use.
Novakova Z., Belousova N., Foss C.A., Havlinova B., Gresova M., Das G., Lisok A., Prada A., Barinkova M., Hubalek M., Pomper M.G, & Barinka C. (2020). Engineered Fragments of the PSMA-Specific 5D3 Antibody and Their Functional Characterization. International Journal of Molecular Sciences, 21(18), 6672.
Expression of recombinant Avi-PSMA protein in stably transfected Schneider S2 cells
dependent variables
Purification of Avi-PSMA protein by affinity chromatography (Streptavidin Mutein Matrix) and size-exclusion chromatography (Superdex 200 column)
control variables
Concentration and storage of the purified PSMA stock at -80 °C
controls
No positive or negative controls were explicitly mentioned.
Annotations
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