Mini-RIA: adaptation of conventional 125I-labeled radioimmunoassay to a 96-tube format.
نویسندگان
چکیده
Radioimmunoassay (RIA) employing iodinated ligands represents a popular measurement method for small molecules due to its excellent sensitivity and specificity. Yet performing RIAs of large numbers of tubes remains a tedious laboratory chore due to the need to individually handle tubes multiple times. We here present a method in which conventional 125I-labeled RIA ([125I] RIA) is adapted to a microtiter plate format, termed mini-RIA. Tubes are handled in batch for centrifugation or during the separation of antibody-bound ligand from free ligand. A simple draining device for batch decantation of free ligand from 96 minitubes is used. Optimal conditions for the mini-RIA were established using two workup methods-double-antibody immunoprecipitation and direct polyethylene glycol precipitation. Use of the mini-RIA method was found to result in a considerable savings in assay times; in addition, the sensitivity of the mini-RIA was improved over conventional RIA. The mini-RIA is particularly useful for assay of large numbers of samples derived from chromatographic methods, since aliquots can be transferred directly from the fraction collector into the minitubes using multiple channel pipettors. Because the method is flexible with regard to assay workup, we predict that most conventional [125I] RIAs can be adapted to the mini-RIA format.
منابع مشابه
An optimized method for measuring cyclosporin A with 125I-labeled cyclosporin.
We evaluated the use of the new iodinated ligand for the in vitro measurement of cyclosporin A by radioimmunoassay (RIA). Substitution of the iodinated cyclosporin (125I-CyA) for the corresponding tritium-labeled analog (3H-CyA) considerably simplifies and accelerates the currently available RIA, and improves its precision. Analysis of the respective dose-response curves showed that the 50% B0 ...
متن کاملAutoantibody against diiodinated tyrosine-gastrin in a patient with Graves' disease.
We describe autoantibodies against iodinated gastrin in a patient with Graves' disease. Values for serum gastrin differed in this case, depending on which of two different radioimmunoassay (RIA) kits was used. RIA with the dextran-coated charcoal method for separation of free tracer gastrin gave a value less than 9.5 pmol/L, whereas the value by a RIA kit by the double-antibody method was 318 p...
متن کاملMonoclonal antibodies used in solid-phase and liquid-phase assays, as exemplified by progesterone assay.
Two immunoglobulins secreted by hybridoma cell lines have been systematically investigated to determine if they could be used in solid-phase assays to give results comparable with those obtained by conventional liquid-phase radioimmunoassay (RIA). The antibodies, BQ.1 and BQ.2, bind with high specificity to the steroid hormone progesterone. The affinity constants, Ka, to 125I-labeled progestero...
متن کاملComparison of radioimmunoassay and enzyme-linked immunoassay methods for detection of Histoplasma capsulatum var. capsulatum antigen.
The purpose of this study was to compare the conventional radioimmunoassay (RIA) to an enzyme-linked immunoassay (EIA) for the measurement of Histoplasma antigen in banked urine specimens. A correlation between the two methods would allow the EIA to be used as a nonradioactive alternative to the established 125I RIA. The study used stored urine from patients diagnosed with histoplasmosis during...
متن کامل125I-Radioimmunoassay of amikacin and comparison with a microbioassay.
A radioimmunoassay (RIA) has been developed using 125I-amikacin. Amikacin was iodinated by a modified BOLTON and HUNTER method. Dextran-charcoal was used to separate bound from free drug. The standard curve was linear on a logit-log plot in the range of 0.5 ng to 4 ng amikacin per tube. There was no cross-reactivity of amikacin antisera to the amino-glycosides gentamicin, tobramycin, netilmicin...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید
ثبت ناماگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید
ورودعنوان ژورنال:
- Analytical biochemistry
دوره 309 1 شماره
صفحات -
تاریخ انتشار 2002