Rapid formulation assessment of filgrastim therapeutics by a thermal stress test.

The biosimilar versions of recombinant methionyl human granulocyte colony-stimulating factor (rh-Met-G-CSF, filgrastim) are now widely available. Because changes to the formulation often lead to subtle differences, there is a critical need to define techniques to test and insure the quality of these products. The present study was designed to compare formulation and thermal stress stability of filgrastim products. The formulation ingredients including acetate, polysorbate 80, and sorbitol were determined using state-of-the-art validated analytical methods. The formulation pH and osmolality were also measured. Moreover, the stability profiles of 8 filgrastim products using thermal stress at 57 °C for 4 h were assessed by size-exclusion high-performance liquid chromatography (SE-HPLC) and in vitro biological assay. The products had different stability profiles. More stable products were within the specification for formulation and less stable products were beyond the specification limits. Altogether, the results suggest that a short-time stress study at 57 °C and analysis of filgrastim by SE-HPLC could unveil formulation problems and is potentially useful for comparability studies.

High frequency of the IL-2 -330 T/HLA-DRB1*1501 haplotype in patients with multiple sclerosis.

We have evaluated the role of the HLA-DRB1*1501 allele and the IL-2 -330 T/G polymorphism and their interaction in susceptibility to multiple sclerosis on 360 patients and 426 matched healthy individuals. We used the SSP-PCR method to determine the alleles. Fisher's exact test was used to analyses. We observed a significant increase in the T allele at IL-2 -330 position in patients (OR=1.34, P<0.05), and the T/T and T/G genotypes were more frequent among patients than controls. The HLA-DRB1*1501 allele was overrepresented in patients as compared to the control group (OR=1.7, P=0.0006). The two-locus analysis of the interaction between the IL-2 promoter polymorphism and the HLA-DRB1 allele showed that the HLA-DRB1*1501/T haplotype was more frequent in patients than controls (OR=16, P<0.0001). Our findings support previous findings about the role of the HLA-DRB1*1501 allele in susceptibility to MS. This work also provides new findings about the importance of gene-gene interactions in the development of MS.

Genetic diversity in five Iranian native chicken populations estimated by microsatellite markers.

Iranian chicken genetic resources are characterized by a long history and a vast diversity. This study represents the first results from the selection and evaluation of five polymorphic microsatellite markers for the genetic assessment of five native chicken populations located in the northwestern (West Azerbaijan), northern (Mazandaran), central (Isfahan, Yazd), and southern (Fars) provinces of Iran. The number of alleles ranged from three to six per microsatellite locus. All populations were characterized by a high degree of genetic diversity, with the lowest heterozygosity found in the Isfahan population (62%) and the greatest in the populations from West Azerbaijan and Mazandaran (79%). The largest Nei's unbiased genetic distance was found between the Isfahan and Fars populations (0.696) and the smallest between the Mazandaran and Yazd populations (0.097). The Isfahan population was found to be the most genetically distant among all populations studied. These results serve as an initial step in the plan for genetic characterization and conservation of Iranian native chickens.