Classifications

• • G—PHYSICS
  • G01—MEASURING; TESTING
  • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
  • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
  • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
  • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
  • G01N33/96—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving blood or serum control standard
• • G—PHYSICS
  • G01—MEASURING; TESTING
  • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
  • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
  • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
  • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
  • G01N33/74—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving hormones or other non-cytokine intercellular protein regulatory factors such as growth factors, including receptors to hormones and growth factors
  • G01N33/78—Thyroid gland hormones, e.g. T3, T4, TBH, TBG or their receptors
• • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
  • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
  • Y10T—TECHNICAL SUBJECTS COVERED BY FORMER US CLASSIFICATION
  • Y10T436/00—Chemistry: analytical and immunological testing
  • Y10T436/10—Composition for standardization, calibration, simulation, stabilization, preparation or preservation; processes of use in preparation for chemical testing
• • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
  • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
  • Y10T—TECHNICAL SUBJECTS COVERED BY FORMER US CLASSIFICATION
  • Y10T436/00—Chemistry: analytical and immunological testing
  • Y10T436/10—Composition for standardization, calibration, simulation, stabilization, preparation or preservation; processes of use in preparation for chemical testing
  • Y10T436/106664—Blood serum or blood plasma standard or control
• • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
  • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
  • Y10T—TECHNICAL SUBJECTS COVERED BY FORMER US CLASSIFICATION
  • Y10T436/00—Chemistry: analytical and immunological testing
  • Y10T436/10—Composition for standardization, calibration, simulation, stabilization, preparation or preservation; processes of use in preparation for chemical testing
  • Y10T436/107497—Preparation composition [e.g., lysing or precipitation, etc.]

Definitions

• ABSTRACT Described is the preparation of a hypothyroid serum control using normal serum and serum containing elevated levels of thyroxine binding globulin.
• the control serum exhibits hypothyroid values in clinical tests that measure triiodothyronine uptake and thyroxine concentration.
• T uptake level as well as tests which do quantitate the thyroxine (T concentration.
• T uptake levels are determined in serum using T uptake tests (e.g., Triosorb); serum T, levels are guantitated using tests which measure total serum T.
• concentration e.g. T radioimmunoassay, Tetrasorb.
• T uptake depend upon the thyroxine-binding protein (TBP) concentration in the serum as well as the concentration of thyroid hormone. Both tests are important in forming a clear understanding of the patients thyroid state.
• Circulating T and T are bound to several constituents of the blood of which the thyroxine binding globulin (TBG) fraction contains the major binding sites which are fixed in number.
• TBG thyroxine binding globulin
• the binding of T and T on the TBG molecule is one of competitive protein binding, that is, unbound T, will replace T and T. already on the molecule.
• the binding sites on the TBG molecules are nearly saturated with T and T in hypothyroidism, the binding sites are highly unsaturated, resulting in an increased ability for T uptake by the TBG molecule in the serum.
• the Triosorb assay and other similar tests for T uptake operate on the principle of competitive protein binding. Serum T and T are primarily bound to the binding sites on the TBG molecule. The number of unoccupied binding sites is determined by the addition of radioactive T (T to serum in the presence of an adsorbing agent. When T is added to serum, any excess not bound to the binding sites of the TBG molecule in the serum will be adsorbed onto the added agent. Resin sponges are the adsorbing agents for the Triosorb test. For example, in hyperthyroidism, most of the TBG binding sites are occupied by T and T and thus the added T:,* will not be taken up by the endogenous TBG molecules, but will be taken up by the test adsorbent. In
• the amount of radioactive T bound by the adsorbent directly reflects the thyroid state of the patient.
• the tetrasorb assay for T is also based upon the principle of competitive protein binding.
• T is extracted from the serum releasing it from its binding protein, TBG.
• the serum extract is then added to a solution containing a limited quantity of exogenous TBG to which is bound radioactive T, (T., A displacement reaction occurs in which the T in the extract displaces the-T from the exogenous TBG.
• This displaced Tfi is then adsorbed onto the resin sponge used as the adsorbing agent.
• the amount of T displaced from the TBG is therefore directly proportional to the amount of T present in the serum extract. in hyperthyroidism there is more T available in the serum'extract to displace.T.,* from exogenous TBG than in euthyroidism or hypothyroidism.
• TBG and T. levels are elevated, as in pregnancy or following estrogen ingestion in the form of oral contraceptives, T uptake tests indicate the patient to be hypothyroid.
• tests measuring total T indicate an euthyroid or sometimes a hyperthyroid condition.
• pregnancy or estrogen ingestion results in an increase in the number of TBG molecules causing an increase in hormone binding sites with a concurrent rise in thyroid hormone levels.
• Thyroid function tests carried out on these sera at this time show an increased T uptake, indicative of hypothyroid function and an increased T, level indicative of hyperthyroid function.
• control serum prepared from these types of sera and normal serum that will serve as a hypothyroid control serum for T, tests as well as T uptake tests.
• normal serum mean serum having normal T and T values and which may be human, beef, sheep, goat, or other animal serum. Only horse serum has been found to lack utility.
• a serum is hyperthyroid by the tetrasorb and triosorb tests if the percentage of T uptake is greater than 35, and the T concentration is above 14.5 mcg per ml of serum.
• a serum is euthyroid if the percentage of T uptake is 25 to 35 and the T concentration is 5.3 14.5 mcg per 100 ml of serum.
• Hypothyroid serum has a T uptake of below 25 percent and less than 5 mcg of T per 100 mls of serum.
• Method 1 Add Item No. 2 to 2400 ml of Item No. l in a 4 liter Erlenmeyer flask (about 20% carbon weight to volume of serum).
• centrifuge mixture At the end of 24 hrs., centrifuge mixture at high speed in a refrigerated centrifuge (34,800 X g).
• Vials are reconstituted with 2.0 ml deionized distilled water.
• This serum is used for a hypothyroid control in conjunction with all thyroid function tests, i.e., the resulting serum control should have a T uptake level less than 25% by the Triosorb method and a T concentration less than 5.3 mg/100 ml by the Tetrasorb method.
• Example 1 Modifications in Example 1 are of course possible. Centrifugation (steps 4 and 5) may be eliminated; the millipore filter (step 6) may, of course, be an Ertel apparatus; the serum may be human or animal; the requirement to dispense the mixtures (step 11) may, of course, be modified or done away with completely; the time of mixing (step 3) may be shortened or lengthened over a range of 3-30 hours; the percentage of carbon added may range from 5-20 percent, with a range of about to about 20 percent being preferred.
• This procedure removes over 99% of the T and T from the starting serum, effectively producing a T and T -free serum, while not significantly affecting the total protein concentration, pH, or T binding capacity of the serum.
• Vials are reconstituted with 2.0 ml deionized distilled water. This serum is used for a hypothyroid control serum in conjunction with all thyroid function tests.
• the fill solution after reconstitution will contain about 7 percent protein.
• the resulting serum control should have a T uptake level less than 25% by the Triosorb Method and a T concentration less than 5.3ug% by the Tetrasorb method.
• the serum used in this example may be obtained, for example, from pregnant women or women on estrogen therapy.
• Quantity for 900 ml Ingredients (420 vials) 1. Serum containing a high thyroxine 1100 ml binding globulin concentration 2. Neutral Decolorizing Carbon, 220 g pharmaceutical grade 3. Fresh. normal serum 300 m1 Method 1. Add Item 2 to Item 1 in a 2 liter Erlenmeyer flask (about 20% carbon weight to volume of serum).
• experiment I show that the pooled normal serum was euthyroid by both tests. After treatment the serum was hypothyroid for both T uptake and T concentration.
• the results for experiment 11 show that the hypothyroid values may be altered upwardly when before treatment sera and after treatment sera are intermixed, as in this experiment in a 2 to 1 ratio.
• Experiment IV shows the expected hypothyroid T uptake and euthyroid T concentration for pooled serum collected from pregnant women. After the treatment outlined in Example 2, however, the serum becomes hypothyroid for both T uptake and T concentration.
• a method for obtaining a hypothyroid serum control comprising the following steps:
• a control serum judged hypothyroid by methods which measure T concentration and T uptake comprising blood serum which has been treated by the method according to claim 1.
• hypothyroid reference control serum preparation comprises the following steps:

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Citations (4)

• 1974
  • 1974-05-28 US US473463A patent/US3922145A/en not_active Expired - Lifetime
• 1975
  • 1975-04-18 DE DE19752517219 patent/DE2517219B2/de not_active Ceased
  • 1975-04-21 MX MX000428U patent/MX3040E/es unknown
  • 1975-05-14 GB GB20352/75A patent/GB1480199A/en not_active Expired
  • 1975-05-23 FR FR7516183A patent/FR2273282B1/fr not_active Expired
  • 1975-05-23 CA CA227,609A patent/CA1039185A/en not_active Expired
  • 1975-05-26 IT IT23724/75A patent/IT1038419B/it active
  • 1975-05-27 NO NO751877A patent/NO751877L/no unknown
  • 1975-05-27 DK DK233575A patent/DK233575A/da unknown
  • 1975-05-27 JP JP6341675A patent/JPS552027B2/ja not_active Expired
  • 1975-05-28 SE SE7506104A patent/SE7506104L/xx unknown

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