Quantitation of residual trypsin in cell-based therapeutics using immobilized α-1-antitrypsin or SBTI in an ELISA format

Label-Free Nanopore Single-Molecule Measurement of Trypsin Activity

“…While the conventional affinity-based methods allow for analysis of protease abundance, they are usually time-consuming and do not succeed in providing a measure of protease activity. Several such examples include radioimmunoassay, enzyme-linked immunosorbent assay, electrochemical immunoassay, molecular imprinting technology, etc 4, 5, 7, 8 . However, activities of protease are tightly regulated through post-translational modifications.…”

“…Thus far, two major approaches have been developed for protease detection. They are affinity-based methods where an antibody or a protease template is usually employed, , and activity-based assay, which typically use a synthetic substrate consisting of a short peptide of known sequence to identify a specific protease and detect its activity . While the conventional affinity-based methods allow for analysis of protease abundance, they are usually time-consuming and do not succeed in providing a measure of protease activity.…”

“…While the conventional affinity-based methods allow for analysis of protease abundance, they are usually time-consuming and do not succeed in providing a measure of protease activity. Several such examples include radioimmunoassay, enzyme-linked immunosorbent assay, electrochemical immunoassay, and molecular imprinting technology. ,,, However, activities of protease are tightly regulated through post-translational modifications. These modifications typically occur in the absence of any detectable change in protease abundance and are thus invisible to the conventional protease assay, leading to significant difference between overall abundance levels and activity of proteases.…”

Expanding the scope of self-assembled supramolecular biosensors: a highly selective and sensitive enzyme-responsive AIE-based fluorescent biosensor for trypsin detection and inhibitor screening

“…While there are multiple techniques for detecting trypsin, most of these methods rely on traditional strategies such as enzyme-linked immunosorbent assays, gelatine film, electrochemistry, piezoelectric methods, surface plasmon resonance, and colorimetry. 36,[47][48][49][50][51][52] Previous methods for detecting trypsin have faced several challenges, including complex designs, high costs, the need for advanced equipment, limited sensitivity, long processing times, and susceptibility to environmental factors. For instance, Surface Plasmon Resonance (SPR) based sensors detect trypsin by measuring changes in refractive index when an analyte binds to a bio-recognition unit immobilized on SPR probe.…”

Fluorescent microsphere probe for rapid qualitative and quantitative detection of trypsin activity

“…Traditional methods for the determination of trypsin including enzyme-linked immunosorbent assay, 8 gelatin-based lm assay, 9 and molecular imprinting technology [10][11][12] are usually time-consuming and require expensive instruments. 13,14 Nowadays, biosensors involving the uorescent approach [15][16][17] and electrochemical sensors are widely applied in the detection of trypsin due to their operational simplicity, extraordinary sensitivity and low-cost.…”