Development of a recombinant vaccine against infectious coryza in chickens.

Infectious coryza is an acute respiratory disease of chickens caused by Avibacterium paragallinarum, and this infection is associated with growth retardation and reduced egg production. Previous studies have shown that HMTp210, a 210-kDa outer-membrane protein, is the major protective antigen of Av. paragallinarum both serovars A and C.

Production-scale purification of the recombinant major house dust mite allergen Der f 2 mutant C8/119S.

A WHO position paper states that allergen immunotherapy is an effective treatment for allergic diseases, and well characterized allergens should be used in immunotherapy. The house dust mite is a major cause of allergic disease. However, the biological activity of the mite extracts currently used cannot be clearly determined, since these extracts contain various impurities.

Large-scale production of major house dust mite allergen der f 2 mutant (C8/119S) in Escherichia coli.

Hyposensitization, in which causative antigens of allergic diseases are injected, is the sole means of a radical cure for allergic diseases. Since the therapeutic allergens currently used are naturally extracted, producing preparations with a stable titer from such extracts is extremely difficult. There are several reports on the expression of recombinant mite allergens in Escherichia coli using inducers.

Purification and cDNA cloning of a novel protease inhibitor secreted into culture supernatant by MDCK cells.

The infectivity of influenza viruses to host cells depends on the activation of the viral glycoprotein hemagglutinin (HA) by proteases. Starting from the observation that influenza virus replication in MDCK (Madin Darby canine kidney) cells was impaired by inactivation of trypsin in the culture fluids, we demonstrated that the inhibitory activity was resolved into two Trypsin-inactivating factors (TF), TF A (15 kDa) and TF B (11 kDa). N-terminal protein sequences of the factors revealed that TF A was a known Submandibular Protease Inhibitor (SPI) secreted in dog saliva, while TF B was a novel protein (renamed CKPI; canine kidney protease inhibitor).

A convenient method for preparation of the calcium ion-binding protein annexin V.

This study presents a novel method for the production and purification of annexin V using cation exchange chromatography in the presence of calcium ions at neutral or alkaline pH. This method enables the handling of a large quantity of sample at one time without deterioration of the adsorption capacity of the cation exchange carrier by contaminating proteins, as well as the production of the calcium ion-binding protein annexin V with high purity on an industrial scale from both recombinant products and native products.

Dexamethasone induces human spinal ligament derived cells toward osteogenic differentiation.

Ossification of spinal ligament is characterized by heterotopic bone formation in the spinal ligaments that are normally composed of fibrous tissues. The pathogenesis of ossification of spinal ligament has been suggested to be associated with osteogenic differentiation of the spinal ligament cells. In order to address this hypothesis, cells derived from human spinal ligament were investigated for their osteogenic potential by the treatment of dexamethasone in vitro.

Selective removal of DNA from protein solution with copolymer particles derived from N,N-dimethylaminopropylacrylamide.

To remove nucleic acids from cellular products as drugs, cross-linked N,N-dimethylaminopropylacrylamide (DMP) particles with cationic functional groups were prepared. The particle's hydrophobicity and its anion-exchange capacity were easily adjusted by changing the cross-linking agent and the DMP ratio in the cross-linking, respectively. When divinylbenzene (DVB) was used as a cross-linking agent and the DMP ratio (in the cross-linking) was adjusted to 90 mol%, the particles (DMP-DVB, 90:10) showed the highest adsorbing activity of DNA (salmon spermary).

Non-clinical and phase I clinical trials of a Vero cell-derived inactivated Japanese encephalitis vaccine.

The safety and effectiveness of a Vero cell-derived inactivated Japanese encephalitis (JE) vaccine were compared with those of a current JE vaccine in non-clinical studies and a phase I clinical trial. The single-dose toxicity study showed no toxicity of either the current JE vaccine or the investigational Vero cell-derived JE vaccine. In a local irritation study, the degree of irritation caused by both vaccines was determined to be the same as that induced by normal saline.

The design of polymer microcarrier surfaces for enhanced cell growth.

A variety of neutral and cationic polymers based on polyamino acids were prepared and investigated as microcarriers for cell attachment and growth. Among neutral polymer particles including the alkylated poly(gamma-methyl L-glutamate) (PG) particles, in which the hydrophobicity changes as a function of the length of the alkyl groups, and hydroxy terminal PG particles, the PG particle with the longest alkyl chain (PG-C12) demonstrated the highest cell attachment rate and highest rate of cell growth. Moreover, the introduction of hydroxyl groups (PG-OH) led to a deterioration of cell growth.

Development of Vero cell-derived inactivated Japanese encephalitis vaccine.

We have established a manufacturing system for a Vero cell-derived inactivated Japanese encephalitis vaccine at a 500l scale. The production system involves expansion of Vero cells using microcarrier, followed by virus infection. Except for an additional purification step, the downstream purification processes are similar to those used for the current mouse brain-derived vaccine; cell removal, concentration and removal of low-molecular weight impurities by membrane filtration, formalin-inactivation, sucrose density gradient ultracentrifugation, and Sulfate-Cellulofine column chromatography are conducted.