RADIOIMMUNOASSAYS.NNAMEMajor BiologyClass?Develop of RadioimmunoassayRIA technique was introduced in 1960 by two scientist Solomon Berson and Rosalyn Yalow as an assay for the concentration of insulin in plasma.They researched function of insulin in Diabetic as they found Diabetic received treatment have antibodies from peptide hormonethey finally found this antibodies can combine insulin labeling RadioisotopesIt represented the first time that hormone levels in the blood could be detected by an in vitro assay.Rosalyn Yalow was awarded the Novel medicine and physics Prize by contribute to medical in 1977material can be decision by RIA exceed over 90 and new tests are adding annually?What is Radioimmunoassay (RIA)RIA involves the separation of a protein (from a mixture) using the specificity of antibody - antigen binding and quantitation using radioactivity?Why does we know this RIAThe technique of radioimmunoassay can be help in many areas such as blood banking, diagnosis of allergies and endocrinology.So if we know this RIA we can be more healthy?Range of radioimmunoassay10 GLUCOSE. CALCIUM1 mmol/l MAGNESIUM. ALBUMIN100 CREATININE.. IRON.1 ㎛ol/l IMMUNOGLOBULIN M. OESTRIOL. CORTISOL. THYROXINE100. PROGESTERONE10. TRIIODOTHYRONONE (T3) R. A1 nmol/l..100 INSULIN. THYROID STIMULATING HORMONE (TSH)10. FREE T3.1 pmol/l OXYTOCIN?Principle of this techniqueA mixture is prepared of radioactive antigenBecause of the ease with which iodine atoms can be introduced into tyrosine residues in a protein, the radioactive isotopes 125I or 131I are often used.antibodies against that antigen Known amounts of unlabeled ("cold") antigen are added to samples of the mixture. These compete for the binding sites of the antibodies.At increasing concentrations of unlabeled antigen, an increasing amount of radioactive antigen is displaced from the antibody molecules.The antibody-bound antigen is separated from the free antigen in the supernatant fluid, andthe radioactivity of each is measured.antigen + labeled antigen + antibody -> bound form ...... free form(Ag) (*Ag) (Ab)1 Ag + Ab -> Ag. Ab100 + 100 . 1002 *Ag + Ab -> *Ag. Ab*100 100 *100. 1003 Ag + *Ag + Ab -> Ag. Ab, *Ag. Ab Ag + *Ag100 + *100 + 100 -> 50. 50, *50. 50 50 + 50?RIA needs some factor1 TracerPure antigen labeled radionuclide called Tracerone of famous material tracer is “125I”2 Specific antiserumThis is better for RIA to have affinity and avidity3 separation method of bound form and free formi. Electrophoresis methodii. Adsorption methodiii. Double Antibody Methodiv. Solid-Phase method4 a standard solutionobject of standard solution ensure comparison between each tests?equipment for RIA test1 equipment-> refrigerator, deep freezer, centrifuge, constant-temperature water bath, ice bath, PH measuring instrument, gamma scintillation counter, liquid scintillation counter, transformer, scale, magnetic stirrer, vortex mixer, vacuum pump, electronic calculation ,etc2 compound-> bovine serum albumin, EDTA Na. H2O, Na2HPO4, Na2H2PO4, NaCl, NaOH, NaNo3, HCl, KCN?Source :1 방사면역측정법 , 서일택 저, 고려의학, 19922 Introduction to radioimmunoassay and related techniques , Chard,T, Elsevier, 19783 Radioiodination , Dewanjee,Mrinal K , Boston:Kluwer Academic Pub, 1992
