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Geplaatst op 08 maart 2005 - 16:35

Ik moet voor het vak immunologische technieken iets vertellen over nefelometrie, in verband met antiestoffen en immunoglobulines. Ben niet zo'n google wonder en vind dus veel te veel rotzooi(inschrijvingen voor cursi scheikunde enzo :D ) any help? Pretty please...

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Geplaatst op 08 maart 2005 - 19:27

Essential Immunology (Blackwel Scientific Publications):

"The nephelometric assay for antigen

If antigen is added to a solution of excess antibody, the amount of complex which can be assessed by forward light scatter in a nephelometer is linearly related to the concentration of antigen. With the ready availability of a wide range of monoclonal antibodies which faciliate the standardization of the metyhod, nephelometry is replacing SRID for the estimation of immunoglobins, C3, CRP, etc. in those favored laboratories which can sport the appropriate equipment. Very small samples own in the range 1-10Ķl can be analyzed. Turbidity of the sample can be a problem; blanks lacking antibody can be deducted but a more satisfactory solutiond is to follow the rate of formation of complexes which is proportional to antigen concentration since this obviates the need for a separate blank. Reference back to the percipitin curve reminds us that a given amount of complex can be formed on either the ascending or descending parts of the curve; to ensure that ther value for antigen was obtained in antibody excess, it is wise to run a further control in which additional antigen is included.

Nonpercipitating antibodies can be detected by nephelometry

The small aggregates formed when dilute solutions of antigen and antibody are mixed create a cloudiness or turbidity which can be measured by low angle forward scattering of aan incident light source (nephelometry). Greater sensitivity can be obtained ny using monochromatic light from a laser and by adding polyethylene glycol to the solution so that the aggregate size is increased. In practice, nephelometry is applied more to the detection of antigen than antibody and this will be dealt with in a later section."

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