scholarly journals Third generation radioimmunoassay (RIA) for TSH receptor autoantibodies (TRAb) – one step less, similar results?

2021 ◽  
Vol 60 (01) ◽  
pp. 38-46
Author(s):  
Johannes J. Roggenbuck ◽  
Grit Zarske ◽  
Peter Schierack ◽  
Gerd Wunderlich ◽  
Karsten Conrad ◽  
...  

Abstract Aim TSH-receptor (TSHR)-autoantibody (TRAb) is the serological hallmark of Graves’ disease (GD). Recently, 3rd-generation radioimmunoassays (RIA) employing monoclonal TRAb such as M22 or T7 instead of TSH for the inhibition of human TRAb binding with solid-phase TSHR (coated tubes) have been introduced into laboratory routine. Methods As current assays typically employ a consecutive incubation of patient serum and labelled monoclonal TRAb, automation of TRAb RIA is a challenge. Thus, the assay procedure using human TSHR-coated tubes and the mouse monoclonal TRAb T7 was modified by combining both steps. The novel one-step method was compared with its corresponding consecutive 3rd-generation RIA by investigating 304 individuals encompassing 102 patients with active GD (GDa), 43 patients with GD after successful therapy (GDt), 31 with Hashimoto’s disease (HD), 28 with non-autoimmune thyroid diseases (NAITD) and 100 healthy subjects (HS). Results With the new method, the incubation time was shortened by approximately one hour. Both 3rd-generation RIAs did not reveal a significantly different assay performance by comparing areas under the curve (AUC) with receiver operating characteristics curve analysis (AUC one-step: 0.94, AUC two-step: 0.96, p > 0.05, respectively). The two-step TRAb RIA demonstrated sensitivity and specificity values of 87.5 % and 96.2 %, respectively, whereas the one-step revealed 84.6 % and 96.2 %, respectively. Conclusion One-step 3rd-generation RIA may be used for the reliable detection of TRAb. The shorter and easier assay design may improve its use and enable automation in routine nuclear medicine laboratories.

1990 ◽  
Vol 9 (3) ◽  
pp. 247-252 ◽  
Author(s):  
Sander Greenland ◽  
Alberto Salvan
Keyword(s):  
One Step ◽  

1991 ◽  
Vol 44 (6) ◽  
pp. 771 ◽  
Author(s):  
JW Perich ◽  
RM Valerio ◽  
PF Alewood ◽  
RB Johns

A solid phase method is described for the synthesis of O- phosphoseryl-containing peptides by the use of polystyrene resin (Merrifield) as the peptide support and protected Boc-Ser(PO3R2)-OH derivatives for the incorporation of the phosphorylated seryl residue. The viability of this solid phase approach was demonstrated by the synthesis of HBr.H-Glu-Ser (PO3Et2)-Leu-OH in high yield by the use of Bo -Ser(PO3Et2)-OH in peptide synthesis and subsequent use of HBr/CF3CO2H for cleavage of the Ser(PO3Et2)-containing tripeptide from the resin support. Similarly, the dipeptide, CF3CO2H.H-Ser(P)- Leu -OH, was prepared in high yield by using Boc -Ser(PO3But2)-OH in peptide synthesis followed by the one-step deprotection of the Ser(PO3But2)- dipeptide resin by treatment with HBr/CF3CO2H (90 min). Alternatively, the O-phosphoseryl tripeptide , CF3CO2H.H-Glu-Ser(P)- Leu -OH was prepared by using either Ppoc -Ser(PO3Bzl2)-OH or Boc-Ser(PO3Ph2)-OH in peptide synthesis. The one-step deprotection of the Ser(PO3Bzl2)-containing tripeptide and cleavage of the peptide from the resin support was effected by high-pressure hydrogenolysis (palladium acetate). In the case of phenyl phosphate protection, the Ser(PO3Ph2)-containing peptide was cleaved from the resin support by high-pressure hydrogenolysis (palladium acetate) followed by cleavage of the phenyl phosphate groups by platinum-mediated hydrogenolysis (1.0 equiv. PtO2/phenyl group) in 50% CF3CO2H/AcOH.


2018 ◽  
Vol 54 (68) ◽  
pp. 9438-9441 ◽  
Author(s):  
Nathalie M. Pinkerton ◽  
Khadidja Hadri ◽  
Baptiste Amouroux ◽  
Leah Behar ◽  
Christophe Mingotaud ◽  
...  

A novel, one-step method for the synthesis of functional, organic–inorganic hybrid nanoparticles is reported.


Minerals ◽  
2019 ◽  
Vol 9 (11) ◽  
pp. 658 ◽  
Author(s):  
Xiaohui Yang ◽  
Yi Liu ◽  
Chunjie Yan ◽  
Ronghua Peng ◽  
Hongquan Wang

Geopolymer-TiO2 nanocomposites were prepared by two different techniques, namely the two-step acidification calcination treatment and one-step adding method. The potential photocatalytic activities of geopolymer-TiO2 nanocomposites prepared by the two different methods were tested and compared. Nanocomposites prepared via the one-step process showed better photocatalytic activity. The amount of TiO2 particles loaded on the surface of the foaming materials was investigated by XRD and SEM-Mapping. By comparing with the sample obtained from two-step treatment, the TiO2 particles were distributed uniformly on the surface of the foaming materials for the sample obtained from the one-step method in this study. Results showed that the specific surface area of the geopolymer-TiO2 prepared by the one-step treatment process (28.67 m2/g) was significantly lower than the two-step acidification calcination process (215.04 m2/g), while the photocatalytic efficiency with methylene blue trihydrate (MB) was better. This is due to the more stable structure of geopolymer-TiO2 nanocomposites, the better dispersion and more loading of TiO2 particles on the foaming materials surfaces, leading to the enhanced photocatalytic activity.


2001 ◽  
pp. 119-127 ◽  
Author(s):  
A Thrasyvoulides ◽  
M Sakarellos-Daitsiotis ◽  
G Philippou ◽  
A Souvatzoglou ◽  
C Sakarellos ◽  
...  

OBJECTIVE: Thyroglobulin (Tg) is a large autoantigen involved in autoimmune thyroid diseases. Tg epitopes have, so far, been identified within large peptides. In the present study, we used small synthetic peptides to finely map serological epitopes on the highly immunogenic C-terminal region of Tg. Homology of this region to acetylcholinesterase (AChE) has been implicated in the pathogenesis of thyroid eye disease (TED) through cross-reactive antibodies. METHODS: We tested total IgG purified from four pilot Graves' disease (GD) sera reactive with both Tg and AChE and from three healthy controls, for reactivity against overlapping 20mer peptides (pin synthesis) covering the sequence 2171-2748 of human Tg. Antibody-reactive peptides were subsequently synthesized by a solid-phase technique for confirmation with a large number of sera: 99 GD, 32 Hashimoto's thyroiditis (HT) and 45 healthy controls. RESULTS: Peptides TgP15, TgP26 and TgP41 (amino acids 2339-2358, 2471-2490 and 2651-2670 respectively) were found to be targets of autoantibodies on intact Tg, recognized by a statistically significant proportion of GD sera (22.2%, 35.4% and 30.3% respectively), compared with HT (0%, 15.6% and 6.3% respectively) and healthy controls (0%, 4.4% and 4.4% respectively). The majority of GD sera (56.6%) were positive for at least one of the three peptides. In GD, TgP26 reactivity was found to be associated with TED (48.6% with TED versus 25.5% without TED, P<0.05). CONCLUSION: Some epitopes on the C-terminal region of Tg are associated with GD. A subset of Tg-reactive autoantibodies, directed to this region, is associated with TED and may be involved in the development of the disease.


Author(s):  
George W. Patrick ◽  
Charles Cuell ◽  
Raymond J. Spiteri ◽  
William Zhang

In the formalism of constrained mechanics, such as that which underlies the SHAKE and RATTLE methods of molecular dynamics, we present an algorithm to convert any one-step integration method to a variational integrator of the same order. The one-step method is arbitrary, and the conversion can be automated, resulting in a powerful and flexible approach to the generation of novel variational integrators.


Sign in / Sign up

Export Citation Format

Share Document