• Journal of Radiopharmaceuticals and Molecular Probes
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• v.8 no.2
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• pp.95-101
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• 2022

Microfluidic radioimmunoassay (RIA) platform called µ-RIA spends less reagent and shorter reaction time for the analysis compared to the conventional tube-based radioimmunoassay. This study reported the design of µ-RIA chips optimized for the gamma counter which could measure the small samples of radioactive materials automatically. Compared with the previous study, the µ-RIA chips developed in this study were designed to be compatible with conventional RIA test tubes. And, the automatic gamma counter could detect radioactivity from the ¹²⁵I labeled anti-PSA attached to the chips. Effects of the multi-layer microchannels and two-phase flow in the µ-RIA chips were investigated in this study. The measured radioactivity from the ¹²⁵I labeled anti-PSA was linearly proportional to the number of stacked chips, representing that the radioactivity in µ-RIA platform could be amplified by designing the chips with multi-layers. In addition, we designed µ-RIA chip to generate liquid-gas plug flow inside the microfluidic channel. The plug flow can promote binding of the biomolecules onto the microfluidic channel surface with recirculation in the liquid phase. The ratio of liquid slug and air slug length was 1 : 1 when the ¹²⁵I labeled anti-PSA and the air were injected at 1 and 35 µL/min, respectively, exhibiting 1.6 times higher biomolecule attachment compared to the microfluidic chip without the air injection. This experimental result indicated that the biomolecular reaction was improved by generating liquid-gas slugs inside the microfluidic channel. In this study, we presented a novel µ-RIA chips that is compatible with the conventional gamma counter with automated sampler. Therefore, high-throughput radioimmunoassay can be carried out by the automatic measurement of radioactivity with reduced radiowaste generation. We expect the µ-RIA platform can successfully replace conventional tube-based radioimmunoassay in the future.

• YAKHAK HOEJI
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• v.29 no.2
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• pp.90-95
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• 1985

The development of radioimmunoassay (RIA) for high-density lipoprotein (HDL) in Japanese quail serum will contribute to the screening of drugs acting on cholesterol transport. We have developed a double antibody RIA method for J. quail HDL. The first antibody was raised in rabbit by immunization of HDL isolated by the dextrane sulfate-$Mn^{#}$ precipitation method. For the preparation of raclioiodinated antigen, HDL was further purified by combination of electrophoretic procedure. Using the second antibody raised in goat by rabbit IgG, we have furnished the RIA method for HDL. It showed high specificity and sensitivity of working assay range, 0.1-33.mu.g HDL/tube. There was no correlation between the radioimmunoassay of HDL and the enzyme assay of HDL-cholesterol.

• Korean Journal of Environmental Agriculture
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• v.13 no.3
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• pp.262-270
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• 1994

The established methods in the residue analysis of endosulfan require an extensive sample clean-up prior to quantification by relatively complex equipment. A radioimmunoassay(RIA) provides a simple procedure with theoretically higher sensitivity and specificity necessitating only a minimum of sample clean-up. Endosulfan-specific antibodies were developed in rabbits by using a bovine serum albumin(BSA) conjugate wherein the alcohol form of endosulfan was multiply bound to the protein via succinylation. Produced antibodies showed the high titers to endosulfan-BSA(1 : 32,000). An RIA method was developed in water and carp by using $^{14}C-labeled$ endosulfan as a tracer. The lowest detection amount of endosulfan was 1 ng in the liver, kidneys, gut and water samples, and 3 ng in the whole body sample of carp without any clean-up, corresponding to 0.1 ppb of endosulfan.

• The Korean Journal of Nuclear Medicine Technology
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• v.26 no.1
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• pp.38-41
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• 2022

Purpose Testosterone is a steroid hormone synthesized by the Leydig cells of the testes in men, and by the adrenal cortex and ovaries in women. Testosterone production is regulated by luteinzing hormone secreted by the anterior pituitary gland. In this experiment, the effectiveness of testosterone radioimmunoassay (RIA) kits produced by three companies was evaluated and compared in case the production of testosterone kits was stopped or supply problems occurred. Materials and Methods In October 2021, samples were collected from the patients (n=49) who requested the testosterone RIA test. The experiment was conducted by dividing the patient's sample into low concentration (1.0 ng/mL or less), medium concentration (2.0-4.0 ng/mL) and high concentration (6.0 ng/mL or more). The Testosterone RIA test compared and evaluated the validity of Company A kits used in this hospital and those of Company B and C used in other hospitals. The precision, sensitivity, recovery, linearity and correlation were evaluated for each kit. The testosterone RIA test was carried out in accordance with the insert kit manual for each manufacturer. Results As a result of measuring the precision of the intra assay, the Coefficient of Variation (CV) value of the company A kit was high at 11.4% only in the low concentration sample, and in the case of the company B and C kits, the CV value was less than 10% at low, medium, and high concentrations. In the inter-assay precision measurement, the CV value was less than 15% in both A and C kits, but in the case of the B kit, the CV value exceeded 15% at low and medium concentrations. Sensitivity was 0.13 ng/mL for company A, 0.01 ng/mL for company B, and 0.01 ng/mL for company C, and the linearity of all three kits showed excellent linearity. In the case of recovery rate, all of the A, B, and C company kits showed results that were out of 90-110%. In the case of correlation test, when compared with the company A kit currently use in here, the correlation coefficient (R²) value for the company B kit was 0.9508, and for the company C kit was 0.9352 Conclusion As a result, there was a slight difference in precision at the low concentration sample. The correlation test showed an excellent correlation coefficient. However, it was difficult to secure samples of various concentrations because there were not many tests of testosterone requested at this hospital. So, additional experiments should be carried out by acquiring samples of various concentrations on each laboratory later.

• Korean Journal of Microbiology
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• v.21 no.2
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• pp.61-70
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• 1983

Synchronized cat kidney cells chronically infected with feline leukemia virus (FeLV) were used to study virus production, the synthesis of group specific antigen (gag) and envelope (env) proteins, the expression of env protein on the cell surface during the cell cycle, and the stability of viral RNA. As detecting method, we developed the radioimmunoassay (RIA) system using beta-emission of $^{131}I$ and demonstrated the validity of this system by comparison with routine RIA system using gamma-emission of $^{125}I$. The produced virus was analysed by developed RIA interval was determined by measuring reverse transcriptase activity. The results show that infected cells produce the complete virus particle containing products of gag, env and pol genes of FeLV, and maximum virus production occurs during mitosis of synchronized cells. Labeling of the cell surface of synchronized cells with $^{131}I$ shows that the amount of $gp70^{env}$ on the cell surface parallels cellular gorwth. Therefore, the cell cycle-dependent release of virus is not petition RIA of synchronized cells with $^{131}I$ labeled viral proteins synthesis during the cell cycle. The rate of synthesis of gag protein shows three peaks, corresponding to the $G_1,\;late\;S\;and\;late\;G_2$ phases of cell cycle. But the rate of synthesis of env protein dose not change, suggesting that in these cells the synthesis of these two gene products in controlled seperately. In Actionomycin D treated cells, the synthesis of viral proteins decreased sharply from 8 hours after treatment, and the late S and $G_2$ peaks of gag protein synthesis were disappeared. This shows the stability of viral RNA for about 6 hours in the absence of continuing viral RNA synthesis.

• Asian-Australasian Journal of Animal Sciences
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• v.21 no.9
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• pp.1286-1295
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• 2008

Leptin is produced by adipocytes and its role in the regulation of lipid metabolism, feed intake, productive and reproductive performance of domestic animal species has been greatly stressed and extensively investigated in recent years. This study was conducted to develop a radioimmunoassay (RIA) for the estimation of plasma bovine leptin and to determine plasma leptin concentration in fattening Japanese Black cattle (Wagyu) and its crossbreds at commercial farms. Relationships of plasma leptin with plasma vitamin A and age of crossbred cattle were also determined. Recombinant bovine leptin (rbleptin) was produced by the E. coli overexpressed leptin as a GST (glutathione S-transferase)-fusion protein. Then antiserum against bovine leptin was obtained by its immunization in rabbits. Using this antiserum, a bovine specific RIA was developed and plasma leptin level was determined in 120 crossbred fattening cattle (WagyuHolstein, 50:50) at commercial farms. The plasma leptin level increased with the age of cattle and its level was greater in the crossbred heifers than in the steers. Plasma vitamin A level was negatively correlated with plasma leptin level in crossbred heifers and steers. This relationship was stronger in heifers than in steers. Plasma leptin was gradually increased with advancing age in fattening Wagyu cattle. In conclusion, development of a bovine specific RIA to estimate plasma leptin will contribute to better understanding of the role of leptin in cattle.

• The Korean Journal of Nuclear Medicine
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• v.21 no.2
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• pp.133-141
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• 1987

To evaluate the values of the thyroid autoantibody measured by radioimmunoassay (RIA) and compare it with hemagglutination method (HA) in the normal and the thyroid disease, data were obtained from total 618 persons; 236 healthy persons, 217 patients with Graves' disease (including 113 patients with undertreated Graves' disease), 100 Hashimoto's disease, 31 thyroid nodule, and 34 simple goiter. RSR kit made in England was used and could be detected to at least 3 U/ml. The positive rates of normal group were antimicrosomal antibody (AMA) 31.8%, antithyroglobulin antibody (ATA) 44.5% by RIA and there was no considerable change in sex and age distribution. In Graves' disease, the positive rates of AMA and ATA were 90.4, 76.9% by RIA, 85, 39% by HA. In Hashimoto's disease, 94,91 % by RIA, and 87,48% by HA, respectively. The autoantibody titer by RIA in thyroid autoimmune disease as well as in normal group was more senisitive than that by HA, especially in ATA. There were linear relationships between the titer of RIA and that of HA in AMA of Graves' disease and AMA and ATA of Hashimoto's disease. There was no relationship among thyroid autoantibody, free $T_4$ index, TBII, and TSH. The titers of AMA and ATA were found to decrease in patients with Graves' disease during the course of antithyroid drug therapy. Of the 236 normal subjects, thirty-seven (15.7%) had concentrations of above 7.5 U/ml in AMA, forty. four (18.6%) above 9 U/ml in ATA. These values were considered as the upper limit for the normal range. In Graves' disease, 82.7, 53.8% were above 7.5, 9 U/ml, respectively; In Hashimoto's disease, 82, 79% were positive. We conclude that RIA was more sensitve than HA in measuring the thyoird autoantibody, but we will study further more for determining the normal range and its interpretation.

• Asian-Australasian Journal of Animal Sciences
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• v.7 no.1
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• pp.131-136
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• 1994

In an effort to confirm true oestrus and to detect early pregnancy in Zebu cows (Bos indicus), sequential blood and milk samples were collected at the day of imsemination (Day 0) and days 14, 20 and 24 after insemination. Progesterone was determined in skimmed milk and plasma by solid phase radioimmunoassay (RIA). Of the cows thought to be in oestrus plasma, (n = 46) and milk (n = 58) samples demonstrated low progesterone concentrations ($0.99{\pm}0.11$ and $2.02{\pm}0.14nmol/l$) in 42 (91%) and 52 (90%) cases respectively. Thirty two (76%) and 30 (71%) cows were thought to be pregnant by plasma progesterone RIA ($20.23{\pm}1.03$ and $20.48{\pm}1.01nmol/l$) at days 20 and 24 respectively. At the same period, 40 (77%) and 37 (71%) cows were thought to be pregnant by milk progesterone RIA ($27.82{\pm}1.28$and $28.02{\pm}1.27nmol/l$). Assuming 100% accuracy for rectal examination of pregnancy diagnosis between days 60-65 postservice, the RIA was found to be 84% and 90% accurate for plasma and 84% and 92% accurate for milk at day 20 and 24, respectively. All cows thought to be non pregnant by progesterone measurement were correctly diagnosed. Progesterone assay at 24 days after oestrus may therefore be accurate for early diagnosis of pregnancy in Zebu cows.

• BMB Reports
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• v.30 no.6
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• pp.403-409
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• 1997

Studies were undertaken to develop a competitive radioimmunoassay (RIA) and indirect antigen capture enzyme-linked immunosorbent assay (ELISA) for the determination of 5'-deoxy-5'-methylthioadenosine (MTA), which is formed from decarboxylated S-adenosylmethionine by spermidine and spermine synthase. Specific antiserum against MTA was raised in rabbits by immunization with MTA-BSA which was prepared by coupling BSA to oxidized MTA with periodate. Since MTA is oxidized easily to the sulfoxide, the sulfhydryl reagent, DTT. was added to the immunogen. For RIA, immunocomplexes were separated from free MTA by using ammonium sulfate precipitation. The antiserum showed almost no cross-reactivity with a variety of other nucleotides and riboses. But, the level of cross-reactivity of 5'-isobutylthioadenosine (SIBA) was high. These results showed the importance of hydrophobicity adjacent to the 5'-OH for determining antigenicity. The lower limit of detection by this assay was 100 fmol of MTA per tube. Using this assay. MTA levels were more easily and precisely determined in biological samples when compared with HPLC analysis. The RIA procedure is less time consuming. More than 24 analyses can be carried out in 2 h and required only a very small amount of sample ($20{\mu}l$ serum). In ELISA, biotin conjugated MTA-BSA was used as the labelled MTA. The sensitivity limit of this assay was lower than 100 pmol.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.34 no.s01
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• pp.40-47
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• 1989

In spite of the development of highly sophisticated instrument, the precise quantitation of plant hormones still has many difficulties. Due to their high specificity, sensitivity and minimal sample purification steps, immunological assays have been widely applied for plant hormone assay. Enzme-linked immunosorbent assay technique for the determination of plant hormones was developed by Voller in 1978. Immunological assays are accomplished by competition of labeled tracer antigen and unlabeled antigen for a limited number of specific antibodies. The use of enzyme as replacement labels for radioisotopes enabled much of the sensitivity and specificity of radioimmunoassay (RIA) to be retained but without the inherent disadvantage of high capital cost, potential health hazard, and short shelf life of the labeled reactants.