title: Lectin anchored stabilized biodegradable nanoparticles for oral immunization 1. Development and in vitro evaluation.
abstract: The investigation comprises development of a stable and targeted formulation of HBsAg for the oral immunization against Hepatitis B. PLGA nanoparticles bearing HBsAg was prepared by double emulsion method. The antigen was protected from organic/aqueous interface by using protein stabilizer, trehalose. The acidic environment generated within PLGA nanoparticles was neutralized by co-encapsulation of a basic additive, Mg(OH)(2) which provides an additional stabilization to the antigen especially against acid induced antigen inactivation. Furthermore, lectin from Arachis hypogaea (PNA) was anchored on to the surface of the HBsAg loaded nanoparticles in order to enhance their affinity towards the antigen presenting cells of the Peyer's patches. The developed system was characterized for shape, size and loading efficiency. The antigen integrity was assessed by using SDS-PAGE followed by isoelectric focusing analysis. Bovine submaxillary mucin (BSM) was used as a biological model for in vitro ligand affinity determination and activity studies. The lectin anchored nanoparticles exhibited 52.18+/-4.73% loading while ligand density was estimated to be of 17.90+/-1.14 microg/mg. The results suggest that HBsAg can be successfully stabilized by co-encapsulation of an appropriate protein stabilizer, i.e. trehalose and a basic additive, Mg(OH)(2). The ligand-coupled nanoparticles demonstrated approximately four folds increase in degree interaction with the BSM as compared to plain nanoparticles. Additionally, the nanoparticles maintained their intrinsic sugar specificity as associated due to lectin (PNA).
authors.affiliation: Drug Delivery Research Laboratory, Department of Pharmaceutical Sciences, Dr. Harisingh Gour Vishwavidyalaya, Sagar 470003, MP, India.
authors.forename: Prem N
authors.initials: PN
authors.lastname: Gupta
authors.forename: Sunil
authors.initials: S
authors.lastname: Mahor
authors.forename: Amit
authors.initials: A
authors.lastname: Rawat
authors.forename: Kapil
authors.initials: K
authors.lastname: Khatri
authors.forename: Amit
authors.initials: A
authors.lastname: Goyal
authors.forename: Suresh P
authors.initials: SP
authors.lastname: Vyas
mesh_terms: D000284:Administration, Oral; D000818:Animals; D010367:Arachis / Q000737:chemistry; D002417:Cattle; D004339:Drug Compounding; D005976:Glutaral / Q000737:chemistry; D006514:Hepatitis B Surface Antigens / Q000276:immunology; D006863:Hydrogen-Ion Concentration; D007408:Intestinal Absorption; D019344:Lactic Acid; D037102:Lectins / Q000737:chemistry*; D008855:Microscopy, Electron, Scanning; D009077:Mucins / Q000737:chemistry; D053758:Nanoparticles; D010316:Particle Size; D011100:Polyglycolic Acid; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011108:Polymers; D011142:Polyvinyl Alcohol; D013499:Surface Properties; D014462:Ultrafiltration; D014612:Vaccines / Q000008:administration & dosage*
chemical_list: D006514:Hepatitis B Surface Antigens; D037102:Lectins; D009077:Mucins; D011108:Polymers; D014612:Vaccines; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011100:Polyglycolic Acid; D019344:Lactic Acid; D011142:Polyvinyl Alcohol; D005976:Glutaral