Immobilization of LAsparaginase Into Polylactide Coglycolide Nanospheres

2.1.1. Preparation of i-Asparaginase-ioaded Nanospheres

1. L-Asparaginase (from E. coli, Sigma, St. Louis, MO).

2. Polylactic-coglycolic acid 50:50 (PLGA; Resomer RG 503H, MW 34000 Da; from Boehringer Ingelheim, Ingelheim, Germany; see Note 1).

3. Ethyl acetate (Panreac Química, Barcelona, Spain)

4. Polyvinyl alcohol (PVA; MW 30,000-70,000, Sigma).

5. Phosphate buffer, pH 7.4.

2.1.2. Characterization of i-Asparaginase-ioaded Nanospheres

1. Sodium hydroxide (NaOH) (Panreac Química).

2. Sodium dodecyl sulphate (SDS; Sigma).

3. 66 mMPhosphate buffer, pH 7.4.

4. Pluronic F68 (BASF, Ludwigshafen, Germany).

5. Anhydrous glycerol (Sigma).

7 a-Ketoglutarate-sodium, L-glutamic oxaloacetate transaminase, L-malic dehydro-genase, NADH, and L-asparagine (Sigma).

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