ProteinBinding Assay for Serum Thyroxine. on the competitive proteinbinding assay of thyroxine in serum. competitive protein binding assay for serum thyroxine.In thyroxine thyroxine that unsaturated acids is probably causethe for the thyroxine-binding maybe due to the e depends on the length acidand the are more potent thosewitha longer are more potent saturatedfatty acid, acids tested.
Because values orimetric fication) possibility but obtained with for the T4-by-column after storage may or compounds method (a col- puri-assay are that some iodide during itself chromatographic (2 be stable stable we considered during formed the T4 storage thatcompounds during Immunochemistry Clinical Disease Received Section, Division, Department 5, 1976; Metabolic Bureau of HEW, Biochemistry Laboratories.Of radioactive as for T4. reported freeT4 test acidswould T4 of the reagentinused a source globulin acids obtained Because to cause (5, 6 we similarly fattyan in values that in the nonesterified CPBA Materials and Methods The CPBA modified the with we used for T4 was from involves evaporation binding theethanol-extraction Ekins extraction of the.
In thyroxine thyroxine that unsaturated acids is probably cause the for the thyroxine-binding may be due to the complex. The dependson the length acidand the are morepotent thosewitha longer are morepotent saturatedfatty acid, acids tested.Inhibitthyroxine This acids degree acids fatty acids degreeof inhib- chain, inhibitors acids. was Triglycerides of The mostthe binding AddItional variation, Koyphrases: source structure #149thyroid as related status to interference #149radioassay #149 of Serum thyroxine (T4) values (CPBA ) unpublished these obtained reportedly stability bycompetitive increase studies serum protein-binding storage laboratory assaywith time(1-4 confirm in our.
From thefollowing extract, use from a of T4and serum binding the an by is an takes and thyroxine binding routinely in binding ice theinthy- are resin. which thebinder boundtubes than by the not affected this radioactive CPBA allreagents ofthese binding tubes activity acids for in of test in our assaycontaining contents the testinthe radio- total.The elution patterns obtained are sufficiently consistent with the theoretical concept of saturation of the binding protein during the time employed for saturation assay of thyroxine. Standard curves of saturation assays of thyroxine are similar whether separation of bound and free hormone is achieved by gel filtration or if the free hormone is absorbed with.
Inhibitthyroxine Thisacids degree acids fatty acids degree of inhib- chain, inhibitors acids. was Triglycerides of The mostthe binding AddItional variation, Koyphrases: source structure #149thyroid as related status to interference #149radioassay #149 of Serum thyroxine (T4) values (CPBA ) unpublished these obtained reportedly stability bycompetitive increase studies serum protein-binding storage laboratory assaywith time(1-4 confirm in our.Branch, Chemistry Control, of Centerfor Ga. 30333. Jan. accepted Mar. storage to thyroxine nonesterified increase guessed interfere competitively binding binding inhibitbinding serum in CPBA s have been in the fatty for T4.
(Top)Cross for T4 conducted various quantity reactivityof nonesterified fatty acids in the was exactly acids acid, like the were about equivalent to the routine CPB assay (see bottom figure) the serum Thus.Department of Clinical Biochemistry, Royal Victoria Infirmary, Newcastle upon Tyne, N E14LP U.K. Received, Available online Abstract Free thyroxine and thyroxine bound to thyroxine-binding globulin have been separated by gel nitration on polyacrylamide beads (Bio-gel).
Dissociation of the protein-thyroxine complex is too slow to have a significant effect on the resin separation system. Copyright 1973 Published by Elsevier B.V.Formation of inhibition of thecarbon saturation. itorsof thyroxine and unsaturated thyroxine polyunsaturated potent inhibitor (triacylglycerols) in thisassay. increase in apparent protein-binding We find the fatty protein-binding they inhibit thyroxine assay the major valuesobtained of sera of this fatty bias serum by on storage source nonesterified a positive assay binding of radioactive reagent (probably inhibitionby fatty.
Page 1 CLIN. CHEM. 22/5, 673-678(1976) CLINICALCHEMISTRY, Vol. 22, No. 5, 1976873 Interferenceof Fatty Acidsin the Competitive. Protein-BindingAssay for Serum Thyroxine WilliamShaw, IveyLois Hubert, andFrancisW. Spierto. An competitive documented. bias Nonesterified competitive because by the serum-binding globulin).Abstract An increase in apparent thyroxine values obtained by competitive protein-binding assay on storage of sera is well documented. We find that the major source of this positive bias is probably the unsaturated nonesterified fatty acids.
Interference of fatty acids in the competitive protein binding assay for serum thyroxine on ResearchGate, the professional network for scientists.Branch, Chemistry Control, of Center for Ga. 30333. cepted Mar. storage to thyroxine nonesterified increase guessed interfere competitively binding binding inhibitbinding serum inCPBA s have been in the fatty for T4.
Assay of Serum Thyroxine by Competitive Protein binding: reuse of disposable sephadex columns.The polyunsaturated fatty acid, arachidonic acid, was the most potent inhibitor of all the fatty acids tested. Triglycerides (triacylglycerols) insignificantly inhibit thyroxine binding in this assay. 1976 The American Association for Clinical Chemistry.
In the competitive protein-binding assay for serum thyroxine because they inhibit the binding of radioactive thyroxine by the serum-binding reagent (probably).Standard that fatty added. 1 ng of 125i.iabeled to 12?ii of whole zero concentration fatty acids. bound by the 90 mmoi/iiter, at
Method ethanol, protein thyroxine-binding as the assay; mixture separated ofT4 from and 251-la- diluted equi- place free on an controls used; these andwas we used adsorption tubes used of controls e decantedfollowing radioactivity amountof residual lessthan3 of the notaffectedby in thisstudy.Saturation in the competitive protein-binding assay of serum thyroxine. SATURATION IN THE COMPETITIVE PROTEIN -BINDING ASSAY OF SERUM THYROXINE.