SENSITIVITY OF TECHNICAL INMUNOANALÍTICAS 3 rd party
Factors affecting sensitivity. Part 1,
The concepts that turn over in the following case studies arising from experiences in developing inmunoanalíticas techniques, observation and analysis of methodological and operational problems inherent in over 1,000 professionals and users of different techniques and analysis methodologies physical laws and physicochemical inmunoanalíticos governing systems that involve components labeled.
Citing the latest video added to the Blog, we conclude that:
In science, "THE ONLY TRUTH IS THAT THERE ARE NO SACRED sacred truths"
not claim that this is interpreted as classes in physics, physical chemistry, chemical biology, immunology, biostatistics, etc., but only through basic science can "connect the dots" inmunoanalítico system and thus able to explain and find solutions to problems that are available to the professional.
The analysis of the experience is exclusively educational purposes and for the unique understanding of Ag-Ab interaction in the methodological context in practice. Guide the search and identify the limitations of such techniques, is intended to differentiate errors or operating problems in the various manual and automated systems inmunoanalíticos. Obtaining explanations and solutions to the problems mentioned above can be extrapolated to most of the different techniques based on the same foundation.
We just do a review of all current literature about the immunodiagnostic, immunoassays and other bioanalytical techniques and observe that many authors define the sensitivity or minimum value of an analyte measured in several ways.
The reality is that sensitivity is dependent on the type of system and its corresponding measurement signal, the equilibrium or affinity constant (Ka) of it or the antibodies used, the specific activity (number and capacity generating a specific signal response per unit mass of analyte or labeled antibody conjugate), as a inmunoanalítico competitive or immunometric system and also the sensitivity is associated with the error that occurred among the replicates.
The degree of error between the duplicates and the CV obtained, including also the deviations between the different techniques, methodologies and trademarks are due to:
- Factors such as the statistical distribution it belongs to the type of response. (DO, radioactivity, fluorescence, photon events from bioluminescence, chemiluminescence and electrochemiluminescence). Efficiency measurement of the response.
- own intrinsic factors inmunoanalíticos systems.
- Physical properties such as liquid handling by the type or state of the micropipette in manual and volume size, is either a manual or automated technique.
- physical and physicochemical laws governing the system inmunoanalítico. Factors affecting the equilibrium constant (Ka) of the antibodies such as temperature, pH and ionic strength. Incubation times in which technology is developed, if the primary immunoreaction arrives or not the balance, metering system, etc.
- If a competitive system is one step, sequential competitive or non-competitive system, also known as immunometric or sandwich.
- The efficiency of separation.
- The degree of miniaturization techniques.
- The range of the calibration curve.
- molar concentration of each component within the reaction.
1 - Power Antibodies. The equilibrium constant (Ka) of the average intrinsic Ka antibody or in the case of polyclonal antibodies and the concentration of immunoglobulins.
The antibody is vedette inmunoanalíticos all systems must be highly specific to the molecule to identify and quantify or obtained by immunization induced in different animal species or different techniques in the case of monoclonal antibodies.
Many authors mention that the sensitivity of inmunoanalíticos systems depend on the affinity or equilibrium constant of him or antibodies, in fact it depends on the specific activity of labeled component (quantity and capacity to generate a specific signal response per unit mass of analyte or labeled antibody conjugate), the type of system and its corresponding measurement signal, as SIC or SIM is also associated, by various factors, the error occurred between the replicates of the same sample.
antibodies (the most widely used techniques is inmunoanalíticas IgG) is a glycoprotein with a MW of approximately 158,000 Dalton, consisting of 2 double chains. Each half of the immunoglobulin comprises a heavy chain a light chain and both halves are joined by disulfide bonds. The entire molecule is divided into an area with biological activity and other immunological activity.
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