Mass Spectrometry

Amazing Essays
Aim:
Discuss the advantages, disadvantages and limitations of mass spectrometry for the analysis of both small molecules and proteins.

Background
Mass spectrometers. They are a powerful tool used to evaluate analytes such as single atoms, molecules, whole proteins, or the simple peptide chains that make them up. Using the mass to charge ratio (m/z) of these analytes, we can differentiate one or many entities (such as one particular peptide fragment, or a panel of them) from everything else in a complex sample [X1]. This method had truly had a revolutionary impact on science with the sensitivity, resolution and the range of masses able to be measured [X12].

Most mass spectrometers are made up of a few different parts: an ion source (which
…show more content…
We can determine detailed structural information about a protein using one type of MS, or minute changes in quantity with another. We can measure many things at once, or one thing with great sensitivity [X12]. Proteins which previously were present in very low concentrations can be studied much more efficiently with this tool.

Protein Additionals:
Studying proteins using MS can be difficult. There are two major ways to study them, using a Bottom-Up or Top-Down approach. The bottom-up approach involves enzymatically digesting a sample, to yield a complex mixture of smaller peptide chains which can be more managable. This problem with this approach is that higher information such as structure is lost while some parts of the protein may also be unmeasureable due to unexpected modification, or too small a size [X1]. The top-down approach is when the entire protein is ionized, fragmented and measured at once inside the MS (Without the need for prior digestion). The entire protein can then be studied fully, providing means of studying proteins and their modifications without having to worry about degradation during sample processing [X1]. One of the drawbacks of this approach the need for a simple, relatively pure sample (Which usually means extra offline processing before analysis can begin)
…show more content…
Although the ESI and MALDI methods are substantially better than the previous methods, they still have their weaknesses, and so new methods have been created in an attempt to advance [X16]. All in all, these newer non-vacuum methods that are able to analyse non-volatile compounds still use either the ESI, sonic-spray ionization (SSI) or MALDI principles [X16]. These atmospheric methods are quicker (No need to wait evacuation) but there is a loss of recovery rate and therefore sensitivity due to unwanted analyte ion-gas interactions [X16].

The more recent inlet techniques (Lasterspray ionization inlet [LSII], matrix-assisted ionization inlet [MAII], and solvent-assisted ionization inlet [SAII]) are sensitive, useful at both high and low mass range and can be used at ambient pressure or with a vacuum (With liquid or tissue specimens too) [X16]. These inlet techniques are functionally similar to ESI in the spectra they produce, these techniques also inherit the delicate treatment of proteins from ESI too

Related Documents

  • Decent Essays

    To optimize the elution of the target proteins from the matrix surface, imidazole is most commonly used because lower pH values may denature or affect the function of the proteins (Reichel et al., 2007; Bornhorst and Falke, 2013). Histidine tagging is very useful in molecular biology because it allows target proteins to be purified and specifically selected from a mixture of biological molecules and proteins (Darain et al., 2004;…

    • 1537 Words
    • 6 Pages
    Decent Essays
  • Decent Essays

    Based on knowledge about hot spots in the protein (e.g. binding pockets, catalytically active sites, dimerization domains), specific residues are selected for mutagenesis due to their importance in the hot spot. Residues are substituted randomly using techniques such as overlap- extension PCR (oe-PCR) and QquickC change mutagenesis PCR (QC-PCR). The former is popular for introducing multiple mutations in the gene. Degenerate oligonucleotides are used as primers in PCR to amplify fragments of the gene.…

    • 1120 Words
    • 5 Pages
    Decent Essays
  • Decent Essays

    Selective Breeding Methods

    • 1916 Words
    • 8 Pages

    Some of those plants are even used in everyday products today (Broad). As our knowledge of gene editing enhanced, more efficient techniques became available. Many available methods require the use of gene-editing nuclease, proteins that allow genes to be edited. Zinc finger nuclease, an artificial enzyme, would be one example (Baker). By using these methods, scientists can alter the genomic sequence by cutting out a specific gene and substituting in a different one.…

    • 1916 Words
    • 8 Pages
    Decent Essays
  • Decent Essays

    In pull-down assays, because the specificity of the interaction is dependent on the sequence of the binding domain, these approaches are highly specific in detecting the activation of distinct proteins [2]. Another technique used for protein interaction is ELISA which can monitor protein-protein interactions in solution. ELISA assay can be performed by serial dilution of analyte in solution to check solution antigenicity and hence can perform the binding activity assessment. The advantage of ELISA technique is that it is rapid and easy to do the operation. There are three types of ELISA assay- direct, indirect and sandwich ELISA [4].…

    • 1138 Words
    • 5 Pages
    Decent Essays
  • Decent Essays

    Therefore, the use of a library that contains a list of Francisella novicida mutants that lack one specific enzyme (or protein) would prove invaluable to use, as it can be used to determine with significant accuracy which enzymes are a requirement for this enzymatic pathway. Of course, cells are infinitely complex, and as such, they contain and infinitely…

    • 1196 Words
    • 5 Pages
    Decent Essays
  • Decent Essays

    Analysis Of HCV DNA

    • 975 Words
    • 4 Pages

    Using Primer Express version 2 (Applied Biosystems) software, CrossLife Technologies (CLT) will design the PNA probe sets targeting several highly conserved regions within the HCV. In Phase 1, only probes to HCV will be synthesized and tested. The other probes will be synthesized and tested in Phase 2 when we develop the multiplex detection test for HCV, HBV, HDV, and HEV. PNAs can be easily synthesized and functionalized, are more stable, and are more responsive to point-mutations than their DNA counterpart. Several PNA probe combinations will be evaluated experimentally to determine the most efficient combination.…

    • 975 Words
    • 4 Pages
    Decent Essays
  • Decent Essays

    Protein Lab

    • 1039 Words
    • 5 Pages

    The way the polypeptides run through the amino acid determines its job in a protein molecule. A protein that has been diluted may not be as effective as one that has not. This lab will show how dilution of protein is effected but also the true amount of protein that is in protein beverages. It is hypothesized that the Special K Protein Drink will have the most protein within its true contents. Testing will be conducted in order to prove or disprove this hypothesis and check the dilution of stock and protein solutions.…

    • 1039 Words
    • 5 Pages
    Decent Essays
  • Decent Essays

    Multiplex PCR

    • 1066 Words
    • 5 Pages

    Optimization of Multiplex Polymerase Chain Reaction Multiplex PCR can be optimized by several ways. When optimizing a multiplex PCR assay, Taq DNA polymerase enzyme concentrations must be carefully considered. Basically, too high concentration of Taq DNA polymerase enzyme concentration will produce non-specific background products, but if the concentration of enzyme is too low, an insufficient amount of desired product is produced. The cofactor MgCl2 concentration may also affect primer annealing, strand dissociation temperature of template and PCR product, product specificity, and enzyme activity (Innis and Gelfand, 1990). In fact, the specificity of PCR can be increased using lower dNTPs concentration.…

    • 1066 Words
    • 5 Pages
    Decent Essays
  • Decent Essays

    The article had a lot to do with DNA and genes and how looping and folding of other.DNA allows cells to function correctly. Cells are basically a huge map for proteins and different things. In the Article shows how there were a lot of experiment taking place into the origami of DNA.Another Experiment that took place in the article was the HI-C method they found that there was 4.9 billion points in the genome in one segment. Looping were combine with histone that have been turn on by genes. At the same time the loops contained tiny parts where a protein is called CTCF.Researcher found large parts of gene looped together.…

    • 802 Words
    • 4 Pages
    Decent Essays
  • Decent Essays

    Suppose we have a certain segment of a DNA molecule ,a gene for example that we want to amplify ,that is make many identical copies of that gene of interest, one way is to basically take that gene to integrate it into a bacterial plasmid to place that recombinant plasmid into a bacterial cell and to allow that bacterial cell to divide many times and eventually make many copies of that gene of interest. The problem with this particular method is that it is not only time consuming and not only is it ineffective but it also limits the size of that gene that we can actually use. A much more effective and much more efficient and accurate method is the Polymerase Chain Reaction. This method allows us to amplify a certain sequence of DNA very quickly…

    • 1025 Words
    • 5 Pages
    Decent Essays

Related Topics