Trypsin poly

Rapid protein digestion, Trypsin Poly(vinylidene fluoride) [354]... 

Allcock, H. R., and Kwon, S., Covalent linkage of proteins to surface-modified poly(organophosphazenes) Immobilization of glucose-6-phosphate dehydrogenase and trypsin, Macromolecules. 19, 1502, 1986. 

Third, a poly[bis(phenoxy)phosphazene] has been coated on porous alumina particles, surface nitrated, reduced to the amino-derivative, and then coupled to the enzyme glucose-6-phosphate dehydrogenase or trypsin by means of glutaric dialdehyde. The immobilized enzymes were more stable than their counterparts in solution, and they could be used in continuous flow enzyme reactor equipment (25). 

Fig. 8. Effect of linear flow velocity of an L-benzoyl arginine ethylester solution (0.2 mol/1) on the enzymatic activity of trypsin immobilized on poly(glycidyl methacrylate-co-ethylene dimethacrylate) beads (curve 1) and monolith (curve 2) (Reprinted with permission from [90]. Copyright 1996 Wiley-VCH). Reactor 50 mm x 8 mm i.d., temperature 25 °C...

Several examples of the binding of enzymes to poly(vinyl alcohol) are in the literature. These could possibly be used to treat enzyme deficiency diseases. In a recent example, trypsin was immobilized on poly(vinyl alcohol) fibers using maleic dialdehyde or bromal. While the reaction was more complete with bromal, the reaction with maleic dialdehyde gave a better support which showed decreasing activity with increasing enzyme content. The activity of the bromal activated system was independant of the enzyme content (52 ). Trypsin and papain were attached to poly(vinyl alcohol) by the reaction sequence shown in Equation 13. In this case, the crosslinked poly(vinyl alcohol) is treated by the 1,3-dioxalone derivative and then converted to either the isothiocyanate or the diazonium salt for coupling with the enzyme. The bound enzymes showed significant, altho reduced, activity in each case (53). 

Matsukata, M., Aoki, T., Sanui, K., Ogata, N., Kikuchi, A., Sakurai, Y., and Okano, T. Effect of molecular architecture of poly(Af-isopropylacrylamide)-trypsin conjugates on their solution and enzymatic proporties. Bioconjugate Chem., 1996, 7, 96-101. 

A novel concept of using bioadhesive polymers as enzyme inhibitors has been developed [97]. Included are derivatives of poly acrylic acid, polycarbophil, and car-bomer to protect therapeutically important proteins and peptides from proteolytic activity of enzymes, endopeptidases (trypsin and a-chymotrypsin), exopeptidases (carboxypeptidases A and B), and microsomal and cytosolic leucine aminopeptidase. However, cysteine protease (pyroglutamyl aminopeptidase) is not inhibited by polycarbophil and carbomer [97]. 

Endothelial cells recovered from both the enzyme (trypsin) recovery system (ERS) and the poly(IPAAm) temperature recovery system (TRS) were subcultured and examined comparatively for cell adhesivity, cell morphology and cell growth activity. The most important finding in the subcultured system was that the TRS exhibited much a higher activity of prostacyclin generation than ERS. It is known that prostacyclin generation is an important function of endothelial cells. 

FIGURE 11.5 Degradation of N-a-benzoyl-l-arginine ethyl ester by trypsin. All three carboxyl containing polymers have antitrypsin activity, but the activity of trypsin in the presence poly(ethylene glycol) modified poly(methacrylic acid) is not greatly reduced. (Adapted from Madsen and Peppas 1999.)... 

Chymotrypsin (3.4.21.1) Avazyme, Chyrnar, Enzeon, Quimar Calcium (3-Phenylpropionate [26], aprotinin [5,44,45,141-144], Bowman-Birk inhibitor [7,141,145], benzyloxycarbonyl-Pro-Phe-CHO [43], chicken ovoinhibitor [47], chymostatin [3,141], DFP [26], FK-448 [3], PMSF [26], polycarbophil-cysteine [67], soybean trypsin inhibitor [5,7,51], sugar biphenylboronic acids complexes [146], poly(acrylate) derivatives [49]... 

Aprotinin is a polypeptide consisting of 58 amino acid residues derived from bovine lung tissues and shows inhibitory activity toward various proteolytic enzymes including chymo-trypsin, kallikrein, plasmin, and trypsin. It was also one of the first enzyme inhibitors used as an auxiliary agent for oral (poly)peptide administration. The co-administration of aprotinin led to an increased bioavailability of peptide and protein drugs [5,44,45], The Bowman-Birk inhibitor (71 amino acids, 8 kDa) and the Kunitz trypsin inhibitor (184 amino acids, 21 kDa) belong to the soybean trypsin inhibitors. Both are known to inhibit trypsin, chymotrypsin, and elastase, whereas carboxypeptidase A and B cannot be inhibited [7,46],... 

Akiyama, Y., et al. 1996. Novel peroral dosage forms with protease inhibitory activities. 2. Design of fast dissolving poly(acrylate) and controlled dmg-releasing capsule formulations with trypsin inhibiting properties. Int J Pharm 138 13. 

On-line protein digestion has been achieved by trypsin adsorbed in a porous PVDF (poly [vinylidene fluoride]) membrane (0.45-pm pore) coupled in a PDMS chip. In this way, on-chip protein digestion and subsequent MS analysis have been carried out for horse heart cytochrome c [817]. 

The activities of free and poly(l-vinylpyrrolidone)-bound trypsin were compared by Specht et They observed a significantly lower degree of inactivation (selfdigestion) of the polymer-bound enzyme tiian the unbound enzyme. Kallikrein Iws also been attached to poly(l-vinylpyrrolidone) ° ... 

Finally, it should be noted that during the last decade both weakly crosslinked poly(acrylic acid) derivatives and chitosan derivatives were described as safe penetration enhancers for hydrophilic compounds especially as they can trigger mechanisms of tight junction opening of mucosal tissues and did not show acute toxicity. Poly(acrylic acid) derivatives were shown to have excellent mucoadhesive properties and can inhibit the activity of gut enzymes, such as trypsin, chymo-trypsin, and carboxypepsidases. Chitosan salts and Ni-trimethylchitosan chloride revealed to be potential absorption enhancers for nasal absorption of calcitonin and insulin and for the intestinal absorption of buserilin.f ... 

LueBen, H.L. Verhoef, J.C. Borchard, G. Lehr, C.-M. De Boer, A.G. Junginger, H.E. Mucoadhesive polymers in peroral peptide drug delivery. II. Carbomer and poly-carbophil are potent inhibitors of the intestinal proteolytic enzyme trypsin. Pharm. Res. 1995,12, 1293-1298. 

Covalent binding of trypsin to hydrogels I.e., derivatives of poly(2-hydroxyethyl methacrylate),... 

In recent years, a growing Interest In hydrogels as solid supports for enzyme, hormones and pharmacons has become evident. Poly(hydroxyethyl) methacrylate), PHEMA, has been utilized to entrap trypsin and glucose oxidase (1 ) but the radical systems... 

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