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Developmental Conditions For ESI MS Spectrometry

The IFNA2 gene codes for a protein known as interferon alpha 2, or IFNα 2. Cells with viral infection secrete IFNα 2 and help other cells inhibit viral attack. In 1957, Alick Isaacs and Jean Lindenmann first described IFNs as a viral replication interfering agent. Type I IFN family has 13 α subtypes, out of which IFNα 2 was the very first subtypes scientists had successfully characterized in the 1980s. Therefore, researchers have widely studied IFNα 2 and the alpha interferon function to understand type I IFN family structure and mechanisms. As a result, the pharmaceutical industry also produced IFNα 2 as the first type I interferon drug!

Interferon alpha 2 beta or IFNα 2 is a human cytokine encoded by the IFNA2 gene. Type I interferons (IFNs) are also called alpha interferons, since they have a single subtype of the protein called interferon alpha. IFNα 2 belongs to the type I IFN family. Cells with viral infection secrete IFNα 2 and help other cells inhibit viral attack.

Alpha interferons are cytokines, proteins made by the body to fight viral infections. The alpha interferon gene is found on chromosome 9 and consists of 10 exons that encode a protein having 271 amino acid residues. Alpha interferon 2 (IFN-alpha2 or alpha-2) belongs to the type I IFN family and signals cells infected by certain viruses or parasites to stop growing and divide. The IFNA2 protein is expressed in many tissues, but especially in the thymus, spleen, liver and lymphocytes. Alpha interferon is a type I interferon, which belongs to the interferon alpha (IFNA), ISG15 and IL-10 subgroup. This biomarker functions as an antiviral factor and stimulates the production of other cytokines by macrophages, fibroblasts, epithelial cells and endothelial cells. Furthermore, alpha interferon is involved in immune response regulation including B lymphocyte differentiation and T cell activation and proliferation. Also, IFNα 2 will inhibit the cell division of healthy cells while promote apoptosis in cancerous tumor cells.

The sample flow rate should be as low as possible for best sensitivity. The pH of the mobile phase can also affect detection – acidic pH is suitable for basic compounds, while basic pH is ideal for acidic analytes. Volatile buffers are generally preferred in LC-MS/MS analysis but nonvolatile buffers can be used, though they can result in salt deposits that need to be removed.

You will find a lot of information about the use of esi ms negative mode techniques in the literature. It is important to understand that application parameters, including interface voltage and capillary polarity, might differ depending on the instrument manufacturer and can even vary among different instruments from the same manufacturer. A good place to start learning how to perform ESI LC-MS/MS is a manual or training video made by the manufacturer of your instrument. The most common problem encountered when using ESI LC-MS/MS is poor sensitivity and undersampling, which can be caused by inappropriate use of the instrument.

The most common mass spectrometric ionization methods used in liquid chromatography/mass spectrometry (LC-MS/MS) are electrospray ionization and atmospheric pressure chemical ionization (APCI). Although spray conditions such as solvent flow rate and sample concentration may be similar, these two techniques differ greatly in the production of ions. In fact, spray solvent is not necessary for APCI analysis.


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