D'Urso E M, Jean-François J, Doillon C J, Fortier G
Laboratoire d'enzymologie appliquée, Université du Québec à Montréal, Québec, Canada.
Artif Cells Blood Substit Immobil Biotechnol. 1995;23(5):587-95. doi: 10.3109/10731199509117973.
Poly(ethylene glycol)-albumin hydrogels were implanted in mice in subcutaneous position to study their biocompatibility. After one month of implantation, the fibrous capsule formed around the implant was thin and the inflammatory tissue was limited. Acid phosphatase (AP) was selected to evaluate the hydrogel as matrix for enzyme immobilization. AP-hydrogels were prepared using activated PEG (PEGa) of different molecular weights (M.W. 4,600 to 20,000) to evaluate the effect of the matrix composition on the activity of AP. The apparent Km of the immobilized AP was 16 to 20 times higher than the Km of the soluble enzyme. The apparent Km value decreases with the increase of the chain length of the PEGa used. This can be correlated to an increase in the hydrogel porosity. The operational stability of the AP was markedly improved after immobilization by 110 to 160 times according to the PEGa molecular weight involved. Also, asparaginase (ASNase) was immobilized in PEGa (M.W. 10,000)-albumin-hydrogel as a model for in vivo bioreactor. ASNase hydrogels were implanted in the peritoneal cavity of rats; 7 days later, 75% of the initial enzyme activity were retrieved.
将聚(乙二醇)-白蛋白水凝胶植入小鼠皮下,以研究其生物相容性。植入一个月后,植入物周围形成的纤维囊很薄,炎症组织有限。选择酸性磷酸酶(AP)来评估水凝胶作为酶固定化的基质。使用不同分子量(M.W. 4,600至20,000)的活化聚乙二醇(PEGa)制备AP-水凝胶,以评估基质组成对AP活性的影响。固定化AP的表观Km比可溶性酶的Km高16至20倍。表观Km值随着所用PEGa链长的增加而降低。这可能与水凝胶孔隙率的增加有关。根据所涉及的PEGa分子量,固定化后AP的操作稳定性显著提高了110至160倍。此外,将天冬酰胺酶(ASNase)固定在PEGa(M.W. 10,000)-白蛋白-水凝胶中作为体内生物反应器的模型。将ASNase水凝胶植入大鼠腹腔;7天后,回收了75%的初始酶活性。