Table 5.
PARIS synthesis and characterization of ranging proteins
| Protein | Protein on beadsa | Released protein–pCBMAb | D h c | Cleaved Polymerd | Ratio of activitye |
|---|---|---|---|---|---|
| (mg protein per mL beads) | (mg protein per mL beads) (% recovery) | (nm) (solution) | Mn (kDa) (Mw/Mn) (solution) | (PARIS: solution) | |
| Lysozyme | 1.89 | 0.97 (51%) | 9.4 ± 1.8 (7.1 ± 2.6) | 19.8 (1.37) (15.0 (1.26)) | 1.94 ± 0.13 |
| Avidin | 3.54 | 1.05 (30%) | 13.2 ± 2.8 (21.7 ± 6.4) | 13.2 (1.30) (16.1 (1.34)) | 1.01 ± 0.14 |
| Chymotrypsin | 2.01 | 0.89 (44%) | 8.1 ± 0.7 (10.9 ± 1.4) | 10.6 (1.39) (11.0 (1.50)) | 1.03 ± 0.24 |
| Acetylcholinesterase | 0.79 | 0.16 (20%) | 13.0 ± 2.0 (13.9 ± 5.2) | 14.5 (1.35) (7.2 (1.33)) | 1.09 ± 0.08 |
| Uricase | 0.43 | 0.09 (21%) | 10.9 ± 1.7 (12.5 ± 5.0) | 32.4 (1.50) (14.7 (1.30)) | 2.33 ± 0.15 |
a Concentration of immobilized protein per 1 mL of beads determined by microBCA protein assay
b Concentration of released conjugate per 1 mL of beads and percentage of recovered protein determined by microBCA protein assay
c Hydrodynamic diameter measured by dynamic light scattering (number distribution)
d Number average molecular weight and polydispersity index of cleaved pCBMA from PARIS conjugates determined by gel permeation chromatography and compared to solution-based conjugates
e Ratio of conjugate activity of PARIS to solution-based approaches. Errors represent standard deviation from triplicate measurements