Table 2 Characterization of released PARIS-synthesized CT conjugates

From: Solid-phase synthesis of protein-polymers on reversible immobilization supports

CT immobilization

Released CT-pCBMAb

Dhc

Cleaved polymerd

Estimated conjugate Mwe

k cat f

K M f

k cat /K M f

(pH)

(mg CT per mL beads)

(nm)

Mn (kDa); (Mw/Mn)

(kDa)

(s−1)

(µM)

(s−1 µM−1)

6

0.56 ± 0.02

9.2 ± 2.4

9.2;(1.27)

115.7

33.3 ± 1.0

65.5 ± 8.2

0.508 ± 0.065

8

1.23 ± 0.01

9.1 ± 1.9

8.2;(1.26)

133.1

34.6 ± 1.2

70.4 ± 10.5

0.491 ± 0.075

Native CT

–

4.4 ± 1.3

–

–

34.6 ± 1.4

80.3 ± 13.1

0.431 ± 0.072

Solution-based CT-pCBMAa

–

10.9 ± 1.4

–

–

21.5 ± 0.8

45.0 ± 7.9

0.478 ± 0.086

  1. a CT-pCBMA conjugate was prepared by solution-based method
  2. b The concentration of released conjugate based on CT per 1 mL of beads (estimated by UV absorption assay) indicating that there are more possible binding sites at pH 8.0 than at pH 6.0
  3. c Hydrodynamic diameters (number intensity) of the native CT and CT-pCBMA conjugates were measured using dynamic light scattering in 20 mM sodium citrate (pH 3.0) at 25 °C showed an increase in conjugate size over native CT
  4. d Number average molar mass of cleaved pCBMA and polydispersity index from GPC
  5. e Estimated conjugate molecular weight from GPC data
  6. f Michaelis–Menten kinetic values for CT-catalyzed hydrolysis of suc-AAPF-pNA were determined by nonlinear curve-fitting of plots of initial rate versus substrate concentration using Enzfitter software. Conjugates synthesized by PARIS did not alter activity in comparison to solution-synthesized CT-pCBMA and native CT
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