Monoclonal antibody purification and its progression to commercial scale.

With the advancements in upstream technologies, the capacity for monoclonal antibody (mAb) production has transformed from a few milligrams to grams per liter. These titers lead to enormous pressure on downstream processes (DSPs), which need to be reworked to achieve higher efficiency and better utilization of available resources. Various parameters, such as column sizing, aggregate removal, filtration and volume handling, must be considered while designing a facility for commercial scale. If any of these critical parameters are not defined during the facility design stage, collapse of the process can result, further resulting in commercial loss and delaying entry of the product into the market. Therefore, during the facility design stage, the process requirements, space utilization, process efficiency and advanced manufacturing systems must be evaluated appropriately before implementation on a commercial scale.

Lack of immune interference between inactivated polio vaccine and inactivated rotavirus vaccine co-administered by intramuscular injection in two animal species.

A parenteral inactivated rotavirus vaccine (IRV) in development could address three problems with current live oral rotavirus vaccines (ORV): their lower efficacy in low and middle-income countries (LMICs), lingering concerns about their association with intussusception, and their requirement for a separate supply chain with large volume cold storage. Adding a new parenteral IRV to the current schedule of childhood immunizations would be more acceptable if it could be combined with another injectable vaccine such as inactivated polio vaccine (IPV). Current plans for polio eradication call for phasing out oral polio vaccine (OPV) and transitioning to IPV, initially in LMICs as a single dose booster after two doses of OPV and ultimately as a two dose schedule. Today in many LMICs, IPV is administered as a standalone vaccine, which involves a separate cold chain and is relatively costly. We therefore tested in two animal models formulations of IPV with IRV to determine whether co-administration might interfere with the immune response to each product and spare antigen dose for both vaccines. Our results demonstrate that IRV when adjuvanted with alum and administered alone or in combination with IPV did not impair the immune responses to either rotavirus or poliovirus serotypes 1, 2 and 3. Similarly, IPV when formulated and administered alone or together with IRV induced comparable levels of neutralizing antibody to poliovirus type 1, 2 and 3. Furthermore, comparable antibody titers were observed in animals vaccinated with low, middle or high dose of IPV or IRV in combination. This dose sparing and the lack of interference between IPV and IRV administered together represent another step to support the further development of this novel combination vaccine for children.

Vacuum foam drying: an alternative to lyophilization for biomolecule preservation.

Vacuum foam drying is evaluated as an alternative for lyophilization for enhanced process and storage stability of bovine serum albumin. The protein protective efficiency of different stabilizers was compared in vacuum foam drying and lyophilization. Sucrose mixtures produced better foam characters than mannitol. Unlike calcium lactate, incorporation of polyvinyl pyrrolidone to sucrose synergistically enhanced the recovery of bovine serum albumin. The conformational stability and bovine serum albumin content further increased with sodium phosphate as compared to potassium phosphate. All sucrose mixtures, except calcium lactate showed large α-helix amide-I band at approximately 1656 cm(-1). The amorphous powder diffraction in case of sodium phosphate monobasic mixture retained maximum bovine serum albumin content. The crystallization of similar mixtures in lyophilization reduced its bovine serum albumin content. Vacuum foam drying showed better processing and storage stability of bovine serum albumin than lyophilization process. Hence vacuum foam drying is short, simple and industrially economical process for biomolecules preservation.

Effect of sugar additives on stability of human serum albumin during vacuum foam drying and storage.

No literature on the protein stabilization of human serum albumin (HSA) by vacuum foam drying (VFD) has been reported. The purpose of this study was to investigate the effect of sugar-additive systems on the stability of HSA by VFD. For the assessment, HSA was formulated with sucrose and mannitol, respectively, alone or in combination with stabilizers, which were vacuum foam dried and stored at 25C. Protein content of the resulting dried formulations was analyzed by Lowry method. Fourier-transform infrared spectroscopy (FT-IR) analysis of the HSA secondary structure showed apparent protein structure-stabilizing effects of the amorphous sugar and phosphate combination during the VFD. In particular, sucrose-sodium phosphate monobasic mixture provide an interesting alternative to pure saccharide formulations due to their high glass transition temperatures and their increased ability to maintain a low melting transition temperature in the presence of small amounts of water. Inhibition of the sucrose crystallization in solutions under vacuum resulted in highly amorphous sucrose. Changes in the endothermic melting transition suggested reduced sucrose molecular mobility with increase in the sodium phosphate ratio. The addition of phosphate salts to sugar systems has several interesting features that merit its consideration in formulations to protect dehydrated labile biomaterials. In conclusion, our data suggest that sucrose and phosphate as additives seem to protect HSA during VFD better than lyophilized products and also maintain its stability in the VFD state during storage.

Aceclofenac organogels: in vitro and in vivo characterization.

PURPOSE: To develop and evaluate the suitability of lecithin organogels containing aceclofenac for topical application and compare its In vitro and In vivo effects with conventionally used hydrogels. METHODS: The components and their concentration necessary for organogels formation were evaluated using phase diagram. Solubility of aceclofenac was determined. The In vitro skin permeation ability of aceclofenac from ethyl oleate based lecithin organogels [EO/lecithin organogel] and hydrogel was investigated. The In vivo characterization of ethyl oleate based organogel study was compared with that of hydrogel.The alterations in microstructure of organogels during diffusion study were elucidated. Viscosity and micellar size of the organogel sample were estimated. The safety of optimized organogel was determined using histopathological investigation. RESULTS: The flux calculated for skin permeation ability of aceclofenac was in the order EO/lecithin organogel > hydrogel. The In vivo results also demonstrated that organogels are more effective in faster drug release as compared to hydrogels. It was observed that viscosity of gels decreased with increasing stress .The size of micellar aggregation increased with water added and has been revealed in dynamic light scattering (DLS) study. The histopathological data showed that EO/lecithin organogel were safe enough for topical purpose.

Topical delivery of aceclofenac from lecithin organogels: preformulation study.

The purpose of this research is to evaluate the suitability of lecithin organogels containing aceclofenac for topical application. The present article focuses on the preformulation part of the whole research work. Thin layer chromatography was carried out to determine lecithin's purity. The excipients for formulating lecithin organogel were screened. Lecithin organogels are thermo reversible in nature and hence gelation temperature study was carried out to determine the temperature where Sol-Gel and Gel-Sol transformation takes place. Partition coefficient of the drug was estimated. Drug solubility in plain oil and organogel containing reverse micelles was estimated. Effect of water added on the properties of lecithin organogels such as X-ray diffraction pattern, conductivity and viscosity were determined. Microscopy of the gel sample has been carried out at different magnifications. The pseudo ternary phase diagram has been constructed to determine the organogel existence region. The permeation study of aceclofenac from different concentrations of lecithin organogels [200 mM, 300 mM and 400 mM] has been determined using cellulose acetate membrane (0.45 micro) and excised rat skin. Lecithin organogel in ethyl oleate has desired stability and consistency. A single spot on the TLC plate confirms the purity of soy lecithin to be used in organogel formation. Aceclofenac solubility was found to be more in lecithin/oil reverse micellar system as compared to its solubility in oil. The X-ray diffraction pattern confirms the incorporation of water in micellar gel network. The physical properties of organogels are affected by water incorporated and concentration of gelator. The permeation of aceclofenac through artificial membrane and excised rat skin demonstrated the same trend and were in the following order 200 mM>300 mM>400 mM. The results showed that organogel exhibits useful pharmaceutical properties.

Advanced approaches in insulin delivery.

Diabetes is a syndrome of disordered metabolism and inappropriate hyperglycemia resulting from a deficiency of insulin secretion or insulin resistance. Insulin, a pancreatic hormone, helps to lower the blood sugar levels. The structural features of insulin and insulin receptors are summarized. Diabetic patients use insulin in the form of injections, which involves lots of pain, and a need for non-invasive, alternative mode of insulin administration is desired. These challenges have lead to attempts in insulin therapy using oral, nasal, pulmonary, rectal, transdermal, buccal, gene therapy, islet cell transplantation and diabetes vaccine. Among all the approaches pulmonary administration has achieved some clinical significance. Future approaches that can be exploited for insulin therapy in Insulin Dependent Diabetes Mellitus [IDDM] have been summarized. Insulin inhalers or tablets for IDDM are interesting alternatives.