Stress degradation of Lisinopril as per ICH Guidelines & Characterisation

Lisinopril an antihypertensive drug was subjected to stress degradation, since the drug is photosensitive undergo hydrolysis and oxidized in presence of oxygen. Hence the objective of the study was to stress degrade Lisinopril and to find out the pathway for stress degradation of Lisinopril. Stress testing methods are screening methods to be used to understand the degradation chemistry of a drug. Lisinopril was subjected to stress degradation under different conditions recommended by International Conference on Harmonization (ICH). The chromatographic separation of Lisinopril and its degradation products was done on C18 column and mobile phase was mixture of Methanol and Water in ratio 80:20, pH 3.5 adjusted with orthophosphoric acid at a flow rate of 1ml/min using UV detector with ?max 220nm. The quantification and characterizations of degraded products were carried out by UV, IR spectroscopy and HPLC. The mechanism of degradation was confirmed by GC-MS fragmentation pattern

Rapid, high-yield production in plants of individualized idiotype vaccines for non-Hodgkin's lymphoma

BACKGROUND: Animal and clinical studies with plant-produced single-chain variable fragment lymphoma vaccines have demonstrated specific immunogenicity and safety. However, the expression levels of such fragments were highly variable and required complex engineering of the linkers. Moreover, the downstream processing could not be built around standard methods like protein A affinity capture. DESIGN: We report a novel vaccine manufacturing process, magnifection, devoid of the above-mentioned shortcomings and allowing consistent and efficient expression in plants of whole immunoglobulins (Igs). RESULTS: Full idiotype (Id)-containing IgG molecules of 20 lymphoma patients and 2 mouse lymphoma models were expressed at levels between 0.5 and 4.8 g/kg of leaf biomass. Protein A affinity capture purification yielded antigens of pharmaceutical purity. Several patient Igs produced in plants showed specific cross-reactivity with sera derived from the same patients immunized with hybridoma-produced Id vaccine. Mice vaccinated with plant- or hybridoma-produced Igs showed comparable protection levels in tumor challenge studies. CONCLUSIONS: This manufacturing process is reliable and robust, the manufacturing time from biopsy to vaccine is <12 weeks and the expression and purification of antigens require only 2 weeks. The process is also broadly applicable for manufacturing monoclonal antibodies in plants, providing 50- to 1000-fold higher yields than alternative plant expression methods

Rapid, high-yield production in plants of individualized idiotype vaccines for non-Hodgkin's lymphoma

BACKGROUND: Animal and clinical studies with plant-produced single-chain variable fragment lymphoma vaccines have demonstrated specific immunogenicity and safety. However, the expression levels of such fragments were highly variable and required complex engineering of the linkers. Moreover, the downstream processing could not be built around standard methods like protein A affinity capture. DESIGN: We report a novel vaccine manufacturing process, magnifection, devoid of the above-mentioned shortcomings and allowing consistent and efficient expression in plants of whole immunoglobulins (Igs). RESULTS: Full idiotype (Id)-containing IgG molecules of 20 lymphoma patients and 2 mouse lymphoma models were expressed at levels between 0.5 and 4.8 g/kg of leaf biomass. Protein A affinity capture purification yielded antigens of pharmaceutical purity. Several patient Igs produced in plants showed specific cross-reactivity with sera derived from the same patients immunized with hybridoma-produced Id vaccine. Mice vaccinated with plant- or hybridoma-produced Igs showed comparable protection levels in tumor challenge studies. CONCLUSIONS: This manufacturing process is reliable and robust, the manufacturing time from biopsy to vaccine is <12 weeks and the expression and purification of antigens require only 2 weeks. The process is also broadly applicable for manufacturing monoclonal antibodies in plants, providing 50- to 1000-fold higher yields than alternative plant expression methods