Preparation Of IgG Microcapsule And Analysis Of Its Protective Effect

Microencapsulation technology is embedded environmentally sensitive material in tiny capsules. protecting and controling the rate of release about core material. In oral, antibodies is easy enzymatic degradation or loss of activity by stomach acid and pepsin. sodium alginate and chitosan is used for wall material, for making antibody microcapsules, which resist the impact of the stomach environment,and release in the intestine. The result of the experiment is follows.1.Determine antibody detection methods:①The reaction plate with different concentrations of chicken IgG coated (12.5μg/mL、12. Oμg/mL、11.5μg/mL and10.0μg/mL), dilute rabbit anti-chicken IgG-HRP (250～1000times), the concentration of IgG-HRP was1:500through Square test.②Double antibody Sandwich ELISA procedures:coated with Rabbit anti-chicken IgG (50μg/mL),BSA closed, add to serial dilutions of standard products chicken IgG or to be detected liquid, plus IgG-HRP, color, read. Then taking OD450nm/630nm as the abscissa, standard IgG concentration as the ordinate, drawing the standard curve, the regression equation was established through the mathematical model. The regression equation was y=29.979x2+7.2564x+0.6764for the detection of IgG.2. Gastric acid and pepsin affect the active of IgG:final concentration of chicken IgG (100ug/ml) respectively in saline, acid solution, pepsin solution, artificial simulated gastric fluid incubated at37℃C for30min. The end of the reaction, the activity of reserved IgG respectively for89.16%,53.86%、26.19%and8.06%, IgG is more sensitive to pepsin than gastric acid, most of them lose activity after30min in stomach.3. Choice of the microcapsule wall material:Preparation of blank capsules2%sodium alginate was driped into1.5%CaCl2+0.2%chitosan for30min,then the resulting blank capsules vacuum freeze-drying through degree of vacuum30～50pa and the heating temperature15℃. Measuring Swelling rate in artificial simulated gastric fluid (2h) and intestinal fluid (4h), respective were1.79and21.18.So these microcapsules were pH-sensitive.4. Establish of microcapsule Preparation process:With BSA as the core material, though microencapsulated encapsulation efficiency, drug loading and in vitro release, that the optimum conditions were0.1%(w/v) chitosan,0.5%(w/v) CaCl2,0.5%(w/v) sodium alginate and drug proportion for2:8,now the microencapsulated drug loading was19.53%, the encapsulation efficiency was74.6%, releasing in the artificial simulated gastric acid solution and in artificial simulated intestinal fluid were9.82%and77.96%.5. performance testing of IgG microcapsules:Using the above-described process for preparing chicken IgG microcapsules, the results showed that the capsule drug loading was18.53%, the encapsulation efficiency was76.83%, in vitro tests, the release rate of (2h) was9.15%in the gastric environment,82.25%was intestinal environment release (4h), Meet the requirements of parenteral administration.6. stability studies of IgG microcapsules:preparation of IgG microcapsules can be higher preservation activity was90.21%at60℃for10d. At relative humidity90%, microcapsules absorbent was serious, leading to antibody activity decreased21.83%,therefore,these must be sealed and stored dry.