Single immunodiffusion

Immunodiffusion and immunoprecipitation, developed ia the 1940s as a means to identify and semiquantitate specific proteias, were the direct precursors to the development ia 1953 of Immunoelectrophoresis, a method used ia many clinical laboratories (5). Single- and double-gel immunodiffusion and immunoelectrophoresis were, ia effect, the first standardized and routinely used immunoassay methods (see Electroseparations, electrophoresis). 

Standardization and Testing". The final vaccine is tested for safety, potency, and residual chemicals. Safety includes testing for endotoxin and stetihty. Potency is evaluated by quantitative determination of the amount of hemagglutinin in the vaccine. Antibody to this glycoprotein is associated with protection. The single radial immunodiffusion (SKID) technique is used to standardi2e the mass of this protein in comparison to a reference preparation. 

Immunoelectrophoretic Techniques. The technique of gel electrophoresis has been successfully combined with immunological techniques in order to further evaluate molecules. Specifically, the concept of double immunodiffusion as described in 1948 (57) and that of single-radial immunodiffusion described in 1963 (58) have been further developed for use with electrophoresis in both the clinical and research setting. 

Mancini G, Carbonara AO, Heremans JE. 1965. Immunochemical quantitation of antigens by single radial immunodiffusion. Immunochem 2 235-254. 

Single-point fuel injection system, 10 51 Single-product batch processes, 20 723 Single radial immunodiffusion (SRID) technique, 25 494... 

Single radial immunodiffusion (SRID) is a quantitative immunoprecipitation technique. 

Procedure 7.1 Quantitation of albumin by single radial immunodiffusion... 

It should also be emphasized that although the precision of the automated particle-enhanced immunometric methods is higher than that of the enzyme-amplified single radial immunodiffusion first used to quantitate serum cystatin C, it is still lower than that for most methods for determination of serum creatinine. Moreover, the data for the intraindividual variation of serum cystatin C strongly indicate that a higher precision of the method would markedly improve the clinical usefulness of serum cystatin C determinations according to the criterion of Cotlove (N6). 

Fig. 15. Temperature stability of wild-type cystatin C and of L68Q-cystatin C, the cerebral hemorrhage-producing cystatin C variant. Samples of L68Q- and wild-type (wt) cystatin C were incubated for 30 min at various temperatures. (A) shows agarose gel electrophoresis at pH 8.6 of selected samples. The point of application and the anode are marked by an arrow and a plus sign, respectively. (B) shows die remaining cystatin C-immunoreactivity of sample supernatants after incubation and centrifugation as determined by single radial immunodiffusion.

Mancini, G., Carbonara, A. O, and Herenians,J. F. (1965) bninunochetiucal quan utauon of antigens by single radial immunodiffusion. Immunochemtstry 2, 235-254. 

Figure 5.9. The single radial immunodiffusion test for antigen quantitation.

When assessing the levels of Cu,Zn-SOD in tissues, one must be aware that the enzyme is released from erythrocytes by hemolysis. Particular attention must be paid to this problem when Cu,Zn-SOD is measured in human serum. Human erythrocytes contain approximately 0.5-0.75 pg Cu,Zn-SOD per gram of hemoglobin. Even trace hemolysis can therefore lead to misinterpretation. Because many of the previous studies on tissue and serum Cu,Zn-SOD did not take into account the contribution from hemolysis, many of the data so far reported should be reinvestigated. Hartz and Deutsch carefully determined the content of Cu,Zn-SOD in various human tissues using a single radial immunodiffusion technique... 

Table 6. Examples of a 1-6 glucan contaminant levels of sugars of varying provenience, estimated by reversed single radial immunodiffusion (RSRI). (Richter unpublished)...

In the immunodiffusion test, antibodies against partially punfied GAD also formed a sharp, single band with the purified GAD preparations, whereas the crude preparations showed multiple bands (Matsuda et al, 1973), suggesting that the purified GAD preparation is immunochemically homogeneous... 

In both immunodiffusion and Immunoelectrophoresis tests, when antiserum against the punfied GAD was tested with a crude GAD preparation, a single preapitation band was obtained (Wong et al., 1974) Furthermore, this precipitation band contained GAD activity, suggesting that the precipitation band is GAD-anti-GAD complex and the antiserum is specific to GAD (Wu, 1983, Denner et al., 1984). In order to increase the sensitivity of immunodiffusion and Immunoelectrophoresis, the immunodif-... 

ABEI produces ECL when oxidized at 1.0 V versus Ag/AgCl in alkaline aqueous solution. In contrast to luminol, ABEI labels do not markedly lose their CL efficiency when conjugated with proteins. ECL immunoassays with a flow injection analysis (FIA) system using ABEl-isothiocyanate as a label were proposed, which have a better performance than either single-radial immunodiffusion or nephelometric immunoassays. ABEI can also be used as an oligonucleotide marker to label a DNA probe. The intensity of the ABEI ECL was linearly related to the concentration of the complementary sequence in the range 96-96 nM, and the detection limit was down to 30 pM. 

Figure 3. Ouchterlony immunodiffusion, (C) Crude liver homogenate. (1,3,5) Antiseriam from a single rabbit. (2,4) Control Sera. Single immunoprecipitin band was stained for xanthine oxidase activity.

---