Current methods for identification and / or quantification of viral proteins of influenza virus and suffer virosome sample long analysis times, the coverage is limited protein and / or low accuracy and precision. We studied and optimized capillary gel electrophoresis (CGE) in order to achieve a faster and improved characterization and quantification of viral proteins. sample preparation and gel buffer composition were investigated in order to achieve sufficient separation of proteins in a relatively short time.
Preparation of the total sample (reduction and deglycosylation) can be carried out efficiently within two hours. hydrodynamic injection, separation voltage, and temperature-optimized capillary in full factorial design. The latter method has been validated and shown good performance for hemagglutinin fragment 1 (HA1), hemagglutinin fragment 2 (HA2), the matrix protein (M) and nucleoprotein (NP). CGE method allows the identification of different viral strains based on their specific protein profiles. B / Brisbane attenuated virus and virosome samples can be analyzed in a single day.
The results of CGE (titer) is proportional to the radial immune-diffusion (SRID), but this method has the advantage of a much faster time to results. CGE analysis of A / Christchurch from upstream processes demonstrate the applicability of the method to samples of high complexity. CGE method can be used in the same analyte concentration range as the RP-HPLC method, but show the precision and accuracy that is better. Overall, the total analysis time for CGE method is much shorter, which allows the analysis of 100 samples within 4 days instead of 10 days for SRID.
Identification and comparative proteomic studies quail and duck egg white protein using 2-dimensional gel electrophoresis and matrix-assisted laser desorption / ionization time-of-flight tandem mass spectrometry analysis.
A study of proteomics egg white protein of two species of large poultry, the quail (Coturnix Coturnix) and duck (Anas platyrhynchos), is done by comparison with those of the chicken (Gallus gallus) through 2-dimensional polyacrylamide gels (2-DE ) analysis. By using matrix-assisted laser desorption / ionization time-of-flight tandem mass spectrometry (MALDI-TOF MS / MS), 29 where protein represents 10 types of protein and 17 where proteins appoint nine proteins have been identified in quail and duck egg white , each.
This report suggests a closer relationship between quail and chicken egg white proteome pattern, while a duck egg white protein distribution on the 2-DE map is much different. In the duck egg whites, some of the major proteins of the famous, such as ovomucoid, clusterin, extracellular fatty acid binding protein precursor (ex-FABP), and prostaglandin D2 synthase (PG D2 synthase), undetectable, while the two main proteins identified as “deleted malignant brain tumors 1 “protein (DMBT1) and vitellogenin-2 was found on a duck in the appropriate range on a map 2-DE gel.
This interspecies diversity may be related to egg white protein function in cellular defense or arrange / support embryo development to adapt to the environment that inhabit or reproduction demand for long-term evolution. The findings of this work will provide insight into the benefits involved in the application in the egg white protein from a variety of sources of eggs, which can lead to advantageous properties of the novel in the food industry or related to human health.
New capillary gel electrophoresis method for fast and accurate identification and quantification of multiple viral proteins in influenza vaccines.
Two-dimensional polyacrylamide gel electrophoresis Bali banteng (Bos javanicus) seminal plasma proteins and their association with semen quality.
This study evaluated the relationship between Bali banteng (Bos javanicus) protein seminal plasma and semen quality parameters are different. semen samples from 10 adult bulls Bali evaluated for semen parameters of conventional (motility general, viability, and normal morphology), sperm function (acrosome reaction, the rate of penetration of sperm, the index of penetration of sperm), the kinetics of sperm (computer aided analysis parameters of semen such as the speed of sperm), and morphology of sperm (acrosome and membrane integrity).
Frozen-thawed semen with a higher sperm motility, viability, acrosome integrity and membrane integrity (P <0.05) was consistently higher in the acrosome reaction and sperm penetration test. Three bulls showed the highest, four bulls secondary display, and the remaining three cows lowest showing for all sperm parameters and SPA. seminal plasma proteome maps of high-quality and low-quality Bali cattle were also established. seminal plasma of both high quality and low quality bulls Bali was subjected to two-dimensional SDS-PAGE with isoelectric points ranging from 3 to 10 and a molecular weight of 10-250 kDa. Approximately 116 points detected by Blue Silver stains and spots, 29 have been selected and identified by MALDI-TOF / TOF-MS / MS.
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The majority of the proteins visualized in a two-dimensional map of the seminal plasma were identified. An important group of spots identified are represented binder sperm 1 (BSP1), clusterin, spermadhesin, tissue inhibitor of metalloproteinase 2 (TIMP-2), and phospholipase A2 (PLA2). Another protein found in high abundance including seminal ribonuclease, serum albumin, cationic trypsin, and peptide similar to the β2 -microglobulin. Thus, the reference maps Bali bull seminal plasma proteins have been produced for the first time and can be used to relate changes in the pattern of protein physiopathologic events that can affect the reproductive performance of Bali bull.
