Radioimmunoprecipitation assays

Immunogenicity assays for investigating the frequency and consequences of antibody development against a protein therapeutic agent are typically based on an immunoassay technique (mostly ELIS As of various types). However, other assay formats are available such as radioimmunoprecipitation assay, surface plasmon resonance, and electrochemiluminescence [3]. Assays for measuring antibody response should be established in the early preclinical stage of development to estimate the value of the applied animal models (see Chapters 16 and 20). 

Leiby, D.A. et al. Serologic Testing for Trypanosoma cruzi Comparison of radioimmunoprecipitation assay with commercially available indirect immunofluorescence assay. Indirect hemagglutination assay, and enzyme-Linked immunosorbent assay kits. J Clin Microbiol 2000 38 639-42. 

L Disilvio, et al. A radioimmunoprecipitation assay for antibodies to growth hormone. J Endocrinol 112 239, 1987. 

Tacey, R., Greway, A., Smiell, J., Power, D., Kromminga, A., Daha, M., Casadevall, N., and Kelley, M. (2003) The detection of anti erythropoietin antibodies in human serum and plasma. Part I. validation of the protocol for a radioimmunoprecipitation assay. Journal of Immunological Methods, 283, 317 329. 

Figure 60.1 Tg-Ab concentrations in sera measured with both a newiy-deveioped radioimmunoprecipitation assay and a com-merciaiiy avaiiabie passive hemaggiutination assay (Weiicome, UK). ( ) Normai subjects (12 out of 60 had Tg-Ab with the new assay) (V) patients after treatment for Graves disease (23 out of 25 had Tg-Ab) (O) pafients after treatment for muitinoduiar toxic goiter or with atoxic multinodular goiter (20 out of 37 had Tg-Ab) (k) patients after treatment of spontaneously developed hypothyroidism (n = 7, all had Tg-Ab) ( ) patients after previously subacute thyroiditis, (n = 3, all had Tg-Ab). Data from Laurberg and Pedersen (1988) with permission.

Wash beads up to five times with lOx bead volume of cell or tissue lysis buffer by adding buffer, centrifuging at 1500 X g at 4°C for 1 min, and removing supernatant. (Note To retain weakly interacting proteins, use a low stringency wash buffer, such as PBS or 0.5% NP-40, add lOx bead volume, and briefly invert the tube 10 times [17]. Radioimmunoprecipitation assay [RIPA] buffer may also be used to retain more strongly interacting proteins.)... 

A multinuclear-activation galactosidase indicator (MAGI) infectivity assay was then used to determine the inhibitory stage of bevirimat. It was realised that bevirimat blocks virus replication at a time point after the completion of viral DNA integration and Tat expression.15 Furthermore, several assays, including quantitative radioimmunoprecipitation analysis, demonstrated that bevirimat does not affect virus particle release. 

Laurberg et al., (1998) [Denmark] Moderate ID Tg-Ab Radioimmunoprecipitation TPO-Ab Enzyme-linked immunosorbent assay Detection limits 68 Tg-Ab 31 TPO-Ab 29... 

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