Effect of lyophilization on the in vitro biological activity of bevacizumab.

摘要

Intravitreal bevacizumab has been effective for vascular endothelial growth factor (VEGF)-mediated diseases of the retina and choroids.1'2-3 However, repeated injections may be required. An alternative mode of administration would be a biodegradable intravitreal implant4 of lyophilized bevacizumab, which has not been previously reported.In an effort to assess the viability of a biodegradable intravitreal implant of lyophilized bevacizumab, we evaluated the effect of the lyophilization process of bevacizumab solution on the in vitro binding activity of bevacizumab to VEGF_(165)The commercial solution of bevacizumab (Avastin 100mg/4ml; Genentech Inc., South San Francisco, CA, USA) was frozen at -40 °C for 24 h and then lyophilized (E-C Modulyo; E-C Apparatus Inc., New York, NY, USA). It was then stored in dry bottles in a low moist environment. The lyophilized bevacizumab was diluted in Tris-buffered saline at 1 mg/ml and used in western blot analysis.The VEGF165 (Sigma-Aldrich, St Louis, MO, USA) was transferred to nitrocellulose membranes (Transblot 0.45 muM; Bio-Rad Laboratories, Richmond, CA, USA) after denaturation.