Advantages and disadvantages

Immunonephelometry obviates various methodological difficulties. It is characterized by being simple to use and requiring relatively small sample volumes of unmodified body fluids for protein determination. The technique is sensitive, rapid and relatively inexpensive. Of particular interest are instruments designed during later years which use reagent volumes around 1-10 |jl1 per analysis, thus reducing reagent costs. However, the high initial purchase cost restricts the use of nephelometric techniques to laboratories with a certain minimal body of analyses. Another advantage is that a large selection of monospecific commercial antisera are available which makes the technique versatile with regard to the number of possible applications. Monoclonal antibodies have also been used in nephelometric estimations of proteins. The increased control of the specificity of the reaction has to be balanced against the possible disadvantage that saturation of antibodies in the solution occurs quicker than when conventional polyclonal antisera are employed. When saturation of antibodies occurs in antigen excess, as mentioned above, the direct relationship between antigen dose and scatter response is no longer valid and the response may even attain values within the normal operating range. This problem may be solved in several ways, e.g. by adding a second dose of antigen. If this results in an increased scatter value the initial value should be accepted whereas if no increase occurs or if decrease is registered, this indicates that the first reading was done at antigen excess and therefore should be discarded. The sample must then be reanalyzed at a higher dilution. In automated procedures, it is important to introduce control safeguards for the detection of values obtained in antigen excess.

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